Abstract:

The present invention provides methods of prevention and treatment of
cannabinoid receptor-associated diseases and conditions, wherein the
method includes administering a substituted imidazoheterocyclic compounds
having the structure of formula I or a pharmaceutically acceptable salt,
acid salt, hydrate, solvate or stereoisomer of a compound having the
structure of formula I.
##STR00001##
The cannabinoid receptor-associated diseases and conditions preventable or
treatable by the methods of the present invention include pain,
inflammation and pruritis.

Claims:

1. A method of prophylaxis, treatment or inhibition of a cannabinoid
receptor-associated disease, disorder or condition in a mammalian
subject, the method comprising administering to the subject a compound
having the structure of formula I or a pharmaceutically acceptable salt,
acid salt, hydrate or stereoisomer thereof, wherein the cannabinoid
receptor-associated disease, disorder or condition is pain or an
inflammatory disease, disorder or condition:and wherein formula I is as
follows: ##STR00687## Y is selected from the group consisting of NRa
and N+R1R2 X.sup.-;Z is selected from the group consisting
of a bond, --(CH2)p, --CH═CH--, --C≡C--, --CONH-- and
--CO--;Ra is selected from the group consisting of --H,
C1-C8 alkyl, C3-C6 alkenyl, C3-C6 alkynyl,
aryl, C3-C8 cycloalkyl, C3-C8 cycloalkenyl,
--SO2R3, --COR3, --CONR3R4, --CSNR3R4,
--COOR3 and --(CH2)qheterocyclyl, wherein the alkyl,
cycloalkyl, cycloalkenyl, aryl and heterocyclyl of Ra are each
optionally substituted with one to four substituents independently
selected from the group consisting of halo, --OH, oxo, --NH2,
--NO2, --CN, --COOH, --COR3, --OCF3, --CF3,
C1-C6 alkyl, C1-C4 alkoxy, C3-C8
cycloalkyl, phenyl, trifluoromethoxy and trifluoromethyl;Rb is
selected from the group consisting of C1-C8 alkyl,
C2-C8 alkenyl, aryl, --NR5R6, ##STR00688## wherein
the alkyl, alkenyl and aryl of Rb are each optionally substituted
with one to three substituents independently selected from the group
consisting of C1-C4 alkyl, C2-C4 alkenyl,
C3-C6 cycloalkyl, alkoxy, aryl, 5-, 6-, and 7-membered
heterocyclyl, halo, --OH, --NH2, --CN and --NO2;Rc is
selected from the group consisting of halo, C1-C6 alkyl,
C2-C6 alkenyl, C2-C6 alkynyl, C3-C10
cycloalkyl, C3-C8 cycloalkenyl, C1-C4 alkoxy, aryl,
5-, 6-, 7-, 8-, 9-, and 10-membered heterocyclyl; wherein the
C2-C6 alkenyl, C2-C6 alkynyl, C3-C10
cycloalkyl, C3-C8 cycloalkenyl, aryl, 5-, 6-, 7-, 8-, 9- and
10-membered heterocyclyl of Rc are optionally substituted with one
to five substituents independently selected from the group consisting of
C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl,
C1-C4 haloalkoxy, C3-C6 cycloalkyl, C4-C8
cycloalkenyl, halo, --OH, --NH2, (A)(A')(A'')(A''')aryl,
(A)(A')(A'')(A''')heterocyclyl, NR14R15,
(CH2)pNR14R15, --CN, --NO2, oxo, --COOR14,
SOR16, SO2R16, SO2NR14R15,
NR15SO2R16, COR14, CONR14R15 and
NR15COR16; wherein (A), (A'), (A'') and (A''') are each an
independently selected from the group consisting of --H, halo and
C1-C4 alkyl and each heterocyclyl of
(A)(A')(A'')(A''')heterocyclyl is independently selected from the group
consisting of 5-, 6-, 7-, 8-, 9- and 10-membered heterocyclyl;R1 and
R2 are each independently C1-C4 alkyl;R3 and R4,
when either or both are present, are each independently selected from the
group consisting of --H, C1-C6 alkyl, C3-C6 alkenyl,
C3-C6 alkynyl, C3-C8 cycloalkyl, C3-C8
cycloalkenyl, aryl, 4-, 5-, 6-, 7- and 8-membered heterocyclyl; wherein
the alkyl, alkenyl, alkynyl, cycloalkyl of R3 and R4 are each
independently optionally substituted with one to three substituents
independently selected from the group consisting of C1-C6
alkyl, C1-C6 haloalkyl, C3-C8 cycloalkyl,
C1-C4 alkoxy, C1-C4 acyl, aryl, 5-, 6-, 7-, 8-, 9-
and 10-membered heterocyclyl, --NH2, --NO2, --CN, --OH, --COOH,
oxo, and halo; provided that if Ra is SO2R3, then R3
is not --H; alternatively, R3 and R4 taken together with the
nitrogen atom to which they are bonded form a heterocyclyl selected from
the group consisting of 4-, 5-, 6-, 7- and 8-membered
heterocyclyl;R5 is selected from the group consisting of --H,
C1-C8 alkyl and C1-C4 haloalkyl; wherein the alkyl
and haloalkyl of R5 are optionally substituted with one to four
substituents independently selected from the group consisting of
C1-C4 alkoxy, --OH, --NH2, oxo and --CN;R6 is
selected from the group consisting of --H, --CR10R11R12,
--CR10R11COR13, C1-C8 alkyl, C3-C10
cycloalkyl, aryl, haloaryl, (CH2)q-linked 5-, to 10-membered
heterocyclyl; wherein the alkyl, cycloalkyl, aryl, and heterocyclyl of
R6 are optionally substituted with one to three substituents
independently selected from the group consisting of C1-C4
alkyl, aryl, halo, --OH, C1-C4 alkoxy, C1-C4
hydroxyalkyl, --COR13, CONHCH3, --SO2R11,
--SO2NR8R9, --NH2, --NHCO--(C1-C4 alkyl),
--NHSO2--(C1-C4 alkyl), --CN, --NO2 and 5- to
10-membered heterocyclyl; alternatively, R5 and R6 taken
together with the nitrogen atom to which they are bonded form a
heterocyclyl selected from the group consisting of 5- to 10-membered
heterocyclyl, which heterocyclyl substituent of R6 is optionally
substituted with one to two substituents independently selected from the
group consisting of C1-C4 alkyl, C1-C4 haloalkyl,
COOR1, --CONR1R2, halo, oxo and (CH2)q-linked
5-, to 10-membered heterocyclyl;R7 is selected from the group
consisting of --COR3, --COOR3, --SO2R3, and 5-, 6-
and 7-membered heterocyclyl;R8 and R9 are independently
selected from the group consisting of --H, C1-C6 alkyl,
C1-C6 hydroxyalkyl, C1-C6 alkylaminoalkyl,
C1-C6 alkoxyalkyl, C1-C6 cyanoalkyl, C1-C4
haloalkyl, C1-C4 aminoacyl, C1-C4 alkoxy,
C1-C6 alkyl-NHSO2CH3, C2-C4 alkenyl,
(B)(B')C3-C10 cycloalkyl, aryl, (CH2)q-linked 5- to
10-membered (B)(B')heterocyclyl, halo, --OH, C1-C4 alkoxy,
C1-C4 alkyl, --CONH2, --NH2, --CN and --NO2;
wherein (B) and (B') are each independently --H, --OH, C1-C4
alkyl, C1-C4 hydroxyalkyl or C1-C4 acyl,
alternatively: (i) R8 and R9, taken together with the nitrogen
atom to which they are bonded form a heterocyclyl moiety or an 8- to
10-membered spiro-bicyclic heterocyclyl each of which are optionally
substituted with one to three substituents selected from the group
consisting of C1-C4 alkyl, C1-C4 haloalkyl, halo,
--(CH2)q--OH, oxo, COOR1, SO2CH3,
C1-C4 acyl --(CH2)q--CN and aryl; or (ii) R8 and
R9, taken together with the carbon atom to which they are bonded
form a cycloalkyl which is optionally substituted with one to three
substituents selected from the group consisting of C1-C4 alkyl,
C1-C4 haloalkyl, halo, --OH, oxo and aryl;R10 is selected
from the group consisting of --H and C1-C4 alkyl;R11 is
selected from the group consisting of --H, C1-C8 alkyl,
C2-C6 alkenyl, C2-C4 alkynyl, C3-C10
cycloalkyl, aryl, 5-, 6-, 7-, -8-, 9- and 10-membered
--(CH2)qheterocyclyl; wherein the alkyl, alkenyl, alkynyl,
cycloalkyl, aryl and 5-, 6-, 7-, 8-, 9- and 10-membered heterocyclyl of
R11 are optionally substituted with one to three substituents
independently selected from the group consisting of C1-C4
alkyl, C3-C6 cycloalkyl, haloaryl, aryl, and 5-, 6-, 7-8-, 9-
and 10-membered heterocyclyl, halo, --OH, C1-C4 alkoxy,
--NH2, -guanidino, --CN, --NO2, oxo, --COOR10,
--CONR8R9, --SO2NR8R9, --SR10, --SORT
and --SO2R1;R12 is selected from the group consisting of
--H, C1-C4 alkyl and C1-C4 hydroxyalkyl;R13 is
selected from the group consisting of --OR10 and
--NR8R9;R14 and R15 are each independently selected
from the group consisting of --H, C1-C4 alkyl and aryl;
alternatively, R14 and R15 taken together with the nitrogen
atom to which they are bonded form a heterocyclyl selected from the group
consisting of 5-, 6-, 7-8-, 9- and 10-membered heterocyclyl;R16 is
C1-C4 alkyl or aryl;X.sup.- is an anionic counterion;m is an
integer from 1 to 3; each instance of p is independently an integer from
1 to 6; and each instance of q is independently zero or an integer from 1
to 4; andprovided that when Rc is heterocyclyl, the heterocyclyl is
directly bonded through a carbon atom of a ring of the heterocyclyl.

2. The method according to claim 1, wherein the compound having the
structure of formula I is administered in a pharmaceutical composition
comprising a pharmaceutically acceptable vehicle, diluent or carrier.

3. The method according to claim 1, wherein the disease, disorder or
condition is acute or chronic pain.

4. The method according to claim 3, wherein the acute or chronic pain is
selected from the group consisting of inflammatory pain, visceral pain,
neuropathic pain and post-operative pain.

5. The method according to claim 1, wherein the disease, disorder or
condition is an inflammatory disease, disorder or condition.

7. The method according to claim 6, wherein the cardiovascular
inflammation is due to atherosclerosis, ischemia, restenosis or
vasculitis.

8. The method according to claim 5, wherein the inflammatory disease or
condition is selected from the group consisting of asthma, Sjogren's
syndrome, pulmonary inflammation, chronic airway inflammation and chronic
obstructive pulmonary disease (COPD).

9. The method according to claim 5, wherein the inflammatory disease or
condition is selected from the group consisting of allergy, psoriasis,
psoriatic arthritis, eczema, scleroderma, atopic dermatitis and systemic
lupus erythematosus.

10. The method according to claim 5, wherein the inflammatory disease or
condition is selected from the group consisting of arthritis, synovitis,
osteomyelitis, rheumatoid arthritis, osteoarthritis and ankylosing
spondylitis.

11. The method according to claim 5, wherein the inflammatory disease or
condition is selected from the group consisting of septicemia and septic
shock.

12. The method according to claim 5, wherein the inflammatory disease or
condition is selected from the group consisting of diabetes, glucose
intolerance, insulin resistance and obesity.

13. The method according to claim 5, wherein the inflammatory disease or
condition is selected from the group consisting of colitis, ulcerative
colitis, Crohn's disease, IBD (inflammatory bowel disease) and IBS
(Irritable bowel syndrome).

14. The method according to claim 5, wherein the inflammatory disease or
condition is due to tumor proliferation, tumor metastasis or
transplantation rejection.

15. The method according to claim 1, wherein the mammal is a human.

16. The method according to claim 1, wherein the disease or condition is
migraine.

17. The method according to claim 16, wherein the compound having the
structure of formula I is from the group consisting of compounds
(1)-(607).

Description:

RELATED APPLICATIONS

[0001]This application is a continuation-in-part of U.S. Ser. No.
12/337,683 filed Dec. 18, 2008 which is a continuation-in-part of U.S.
Ser. No. 12/142,846 filed Jun. 20, 2008, now U.S. Pat. No. 7,517,874,
which claims the benefit of U.S. Provisional Application Nos. 60/936,754
filed Jun. 21, 2007; 60/994,422 filed Sep. 19, 2007, and 61/008,395 filed
Dec. 19, 2007, the specifications of each of which are hereby
incorporated by reference in their entireties.

FIELD OF THE INVENTION

[0002]The invention relates to uses of substituted imidazoheterocycles,
and more particularly to substituted tetrahydroimidazo[1,5-a]pyrazine and
substituted tetrahydro-5H-imidazo[1,5-a][1,4]diazepine compounds in the
treatment and prevention of cannabinoid receptor-associated diseases,
disorders and conditions, including pain, inflammation and pruritis.

BACKGROUND OF THE INVENTION

[0003]Classical cannabinoids such as the marijuana-derived compound
Δ9-tetra-hydrocannabinol, (Δ9-THC) exert their
pharmacological effects through interaction with specific members of the
G-protein coupled receptor (GPCR) family. To date, two cannabinoid
receptors have been cloned and characterized: CB1, a receptor found in
the mammalian brain and to a lesser extent in peripheral tissues; and
CB2, a receptor found primarily in the peripheral tissues, particularly
in cells of the immune system. Several endogenous ligands for these
cannabinoid receptors, known as endocannabinoids, have been identified.
For a review see Hanus, L. O., Discovery and isolation of anandamide and
other endocannabinoids, Chem. Biodivers. (2007) 8:1828-41.

[0004]Compounds that are modulators of one or both of the cannabinoid
receptors have been shown to produce a variety of pharmacological effects
that may be of therapeutic benefit in humans (see, for example, Mackie,
K., Cannabinoid receptors as therapeutic targets, Ann. Rev. Pharmacol.
Toxicol. (2006) 46: 101-122; Pertwee, R. G., The therapeutic potential of
drugs that target cannabinoid receptors or modulate the tissue levels or
actions of endocannabinoids, AAPS J. (2005) 7:E625-654). The cannabinoid
receptor modulator can be an agonist, an inverse agonist or a neutral
antagonist, and may interact at the same (orthosteric) site as the
endogenous ligand, or at a different (allosteric) site.

[0005]Activation of the CB1 receptor in the brain is believed to mediate
undesirable psychotropic effects associated with Δ9-THC and
other centrally acting cannabinoid ligands. As a result, there has been
considerable interest in developing compounds that possess high affinity
and selectivity for the CB2 receptor (see for example, Raitio, K. H. et
al., Targeting the Cannabinoid CB2 Receptor: Mutations, Modelling and
Development of selective CB2 ligands, Curr. Med. Chem. (2005) 12:
1217-37). CB2 receptor agonists have shown efficacy in preclinical models
of neuropathic and inflammatory pain and may also find application in
cancer, multiple sclerosis, osteoporosis, Alzheimer's disease, liver
disease and diabetes (Mackie, K.; Ross R A; CB2 cannabinoid receptors:
new vistas, Br. J. Pharmacol. (2008) 153: 177-78 and references cited
therein). There is an ongoing need to identify new CB2 ligands that
exhibit greater receptor selectivity, improved drug-like properties and,
for some indications, restriction to the periphery with low or minimal
effects on the central nervous system (CNS).

SUMMARY OF THE INVENTION

[0006]The present invention provides methods of treating, inhibiting and
preventing cannabinoid-receptor associated diseases, disorders and
conditions by administering an effective amount of a composition that
includes a compound having the structure of formula I or pharmaceutically
acceptable salt, acid salt, hydrate, solvate or stereoisomer or mixture
of stereoisomers of a compound of formula I:

##STR00002##

[0007]Cannabinoid-receptor associated disorders, diseases and conditions
that can be treated, inhibited or prevented by administering an effective
amount of a composition that includes a compound having the structure of
formula I or pharmaceutically acceptable salt, acid salt, hydrate,
solvate or stereoisomer or mixture of stereoisomers of a compound of
formula I include, without limitation, pain and inflammation.

[0008]The pain that can be treated, inhibited or prevented by
administering an effective amount of a composition that includes a
compound having the structure of formula I, can be inflammatory pain,
visceral pain, neuropathic pain or hyperalgesia. Each of these types of
pain can present as acute or chronic pain.

[0009]The inflammation that can be treated, inhibited or prevented by
administering an effective amount of a composition that includes a
compound having the structure of formula I, include inflammatory diseases
and conditions associated with elevated levels of one or more
proinflammatory cytokines, including but not limited to tumor necrosis
factor-alpha (TNF-α), interleukin 1β (IL-1β), interleukin
6 (IL-6), interleukin 8 (IL-8), and granulocyte macrophage-colony
stimulating factor, GM-CSF.

[0010]In the compounds of formula I, Y is NRa or
N+R1R2 X.sup.-, wherein X.sup.- is an anionic counterion;
m is an integer equal to 1, 2 or 3; and Z is a bond or a bivalent linking
group chosen from --(CH2)p, --CH═CH--, --≡C--,
--CONH-- and --CO--; wherein p is an integer from one to six.

[0011]The radical Ra is chosen from hydrogen, alkyl having from one
to eight carbon atoms, alkenyl and alkynyl each having from three to six
carbon atoms; cycloalkyl or cycloalkenyl each having from three to eight
ring carbon atoms; aryl; --SO2R3, --COR3,
--CONR3R4, --CSNR3R4, --COOR3, and
--(CH2)q-heterocyclyl; wherein q is zero or an integer from one
to four. The alkyl, cycloalkyl, cycloalkenyl, aryl and heterocyclyl
moieties of Ra are each optionally substituted with from one to four
groups independently chosen from halo, hydroxyl, oxo, amino, nitro,
cyano, carboxyl, --COR3, trifluoromethoxy, trifluoromethyl, alkyl
having from one to six carbon atoms, alkoxy having from one to four
carbon atoms, cycloalkyl having three to eight ring carbon atoms and
phenyl.

[0012]The radical Rb is bonded through the carbonyl of formula I and
is chosen from alkyl having from one to eight carbon atoms, alkenyl
having from two to eight carbon atoms, aryl, --NR3R6,

##STR00003##

and

##STR00004##

wherein the alkyl, alkenyl and aryl of Rb are each optionally
substituted with one to three substituents independently chosen from
alkyl having from one to four carbon atoms, alkenyl having from two to
four carbon atoms, cycloalkyl having from three to six carbon atoms,
alkoxy having from one to four carbon atoms, aryl, and four-, five-,
six-, seven-, eight- and nine-membered heterocyclyl, halo, hydroxyl,
amino, cyano and nitro.

[0013]The radical Rc is chosen from halo, alkyl having from one to
six carbon atoms, alkenyl having from two to six carbon atoms, alkynyl
having from two to six carbon atoms, cycloalkyl having from three to ten
carbon atoms, cycloalkenyl having from three to eight carbon atoms,
alkoxy having from one to four carbon atoms, aryl, and four-, five-,
six-, seven-, eight- and nine-membered heterocyclyl; wherein the alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, aryl, four-, five-, six-, seven-,
eight- and nine-membered heterocyclyl of Rc are optionally
substituted with one to five substituents independently chosen from
alkyl, alkoxy, haloalkyl, and haloalkoxy each having from one to four
carbon atoms, cycloalkyl having from three to six carbon atoms,
cycloalkenyl having from four to eight carbon atoms, halo, hydroxyl,
amino, (A)(A')(A'')(A''')aryl, (A)(A')(A'')(A''')heterocyclyl,
--NR14R15, --(CH2)pNR14R15, cyano, nitro,
oxo, --COOR14, --SOR16, --SO2R16,
--SO2NR14R15, --NR15SO2R16, --COR14,
--CONR14R15 and --NR15COR16; wherein (A), (A'), (A'')
and (A''') are each an independently chosen from hydrogen, halo and alkyl
having from one to four carbon atoms; and each heterocyclyl of the
(A)(A')(A'')(A''')heterocyclyl is independently chosen from four-, five-,
six-, seven-, eight- and nine-membered heterocyclyl.

[0014]The substituents, R1 and R2 are each independently an
alkyl radical having from one to four carbon atoms.

[0015]The substituents, R3 and R4, when either or both are
present, are each independently chosen from hydrogen, alkyl having from
one to six carbon atoms, alkenyl having from three to six carbon atoms,
alkynyl having from three to six carbon atoms, cycloalkyl having from
three to eight ring carbon atoms, cycloalkenyl having from three to eight
ring carbon atoms, aryl and four-, five-, six-, seven-, eight- and
nine-membered heterocyclyl. The alkyl, alkenyl, alkynyl and cycloalkyl of
R3 and R4 are each optionally substituted with one to three
substituents independently chosen from alkyl having from one to six
carbon atoms, haloalkyl having from one to six carbon atoms, cycloalkyl
having from three to eight ring carbon atoms, alkoxy having from one to
four carbon atoms, acyl having from one to four carbon atoms, aryl,
five-, six-, seven-, eight-, nine- and ten-membered heterocyclyl, amino,
nitro, cyano, hydroxyl, carboxyl, oxo, and halo. Alternatively, R3
and R4 taken together with the nitrogen atom to which they are
bonded form a four-membered, five-membered, six-membered, seven-membered
or eight-membered heterocyclyl moiety.

[0016]The substituent, R5 is chosen from hydrogen, alkyl chain of one
to eight carbon atoms and haloalkyl having from one to four carbon atoms;
wherein the alkyl and haloalkyl are optionally substituted with from one
to four substituents independently chosen from alkoxy having from one to
four carbon atoms, hydroxyl, amino, oxo and cyano.

[0017]The substituent, R6 is chosen from the following: hydrogen,
--CR10R11R12, --CR10R11COR13, alkyl having
from one to eight carbon atoms, cycloalkyl having from three to ten ring
carbon atoms, aryl, haloaryl and (CH2)q-linked heterocyclyl
having a four-, five-, six-, seven-, eight-, nine- or ten-membered
moiety; wherein the alkyl, cycloalkyl, aryl, and heterocyclyl are
optionally substituted with from one to five substituents independently
chosen from alkyl having one to four carbon atoms, aryl, halo, hydroxyl,
amino, -cyano, nitro, alkoxy having one to four carbon atoms,
hydroxyalkyl having one to four carbon atoms, --COR13,
--CONHCH3, --SO2R11, --SO2NR8R9, and five-,
six-, seven-, eight-, nine- and ten-membered heterocyclyl.

[0018]Alternatively, R5 and R6 taken together with the nitrogen
atom to which they are bonded can form a five-, six-, seven- or eight-,
nine- or ten-membered heterocyclyl, which is optionally substituted with
one to two substituents independently chosen from alkyl having from one
to four carbon atoms, haloalkyl having from one to four carbon atoms,
halo, oxo, --CONR1R2 and five-, six-, seven- or eight-, nine-
or ten-membered heterocyclyl.

[0020]The substituents, R8 and R9 are independently chosen from
hydrogen; alkyl, hydroxyalkyl, alkylaminoalkyl, alkoxyalkyl and
cyanoalkyl, each having from one to six carbon atoms; haloalkyl having
from one to four carbon atoms, aminoacyl having from one to four carbon
atoms, alkyl-NHSO2CH3 having from one to six carbon atoms,
alkoxy having from one to four carbon atoms, alkenyl chain having two to
four carbon atoms, cycloalkyl having from three to six ring carbon atoms,
aryl; --(CH2)q-linked five-, six-, seven-, eight- nine- and
ten-membered (B)(B')heterocyclyl, halo, hydroxyl, alkoxy or alkyl having
from one to four carbon atoms, amido, amino, cyano or nitro. The
substituents (B) and (B') are each independently hydrogen, hydroxyl,
alkyl or hydroxyalkyl having from one to four carbon atoms.

[0021]In the first of two alternatives, R8 and R9, taken
together with the nitrogen atom to which they are bonded, form a four-,
five-, six, seven-, eight-, nine or ten-membered heterocyclyl moiety, or
an eight-, nine- or ten-membered spirobicyclic heterocyclyl moiety, which
heterocyclyl moiety is optionally substituted with from one to three
substituents independently chosen from alkyl or haloalkyl having from one
to four carbon atoms, halo, oxo, --(CH2)q--OH,
--(CH2)q--CN, COOR1, SO2CH3, acyl having from
one to four carbon atoms and aryl. In the second of two alternatives,
R8 and R9, taken together with the carbon atom to which they
are bonded, form a carbocycle, which carbocycle is optionally substituted
with from one to three substituents independently chosen from alkyl
having from one to four carbon atoms, halo, hydroxyl, oxo and aryl.

[0022]The substituent, R10 is chosen from hydrogen and alkyl having
from one to eight carbon atoms.

[0023]The substituent, R11 is chosen from: hydrogen, alkyl having
from one to eight carbon atoms, alkenyl having from two to six carbon
atoms, an alkynyl chain having two to four carbon atoms, cycloalkyl
having from three to ten ring carbon atoms, aryl, five-, six-, seven- and
eight-membered monocyclic heterocyclyl and nine-membered and ten-membered
bicyclic heterocyclyl; wherein the alkyl, alkenyl, alkynyl, cycloalkyl,
aryl and heterocyclyl of R11 are optionally substituted with from
one to three substituents independently chosen from alkyl having from one
to four carbon atoms, cycloalkyl having from three to six carbon atoms,
aryl, 5-, 6-, 7- and 8-membered monocyclic heterocyclyl, 9- and
10-membered bicyclic heterocyclyl, halo, hydroxyl, alkoxy having from one
to four carbon atoms, amino, guanidino, cyano, amino, oxo, --COOR10,
--CONR8R9, --SO2NR8R9, --SR10, --SOR1
and --SO2R1.

[0024]The substituent, R12 is chosen from hydrogen, alkyl having from
one to eight carbon atoms, and hydroxyalkyl having from one to six carbon
atoms.

[0025]The substituent, R13 is chosen from --OR10 and
--NR8R9.

[0026]The substituents, R14 and R15 are each independently
hydrogen, alkyl having from one to four carbon atoms or aryl; and
R16 is C1-C4 alkyl or aryl.

[0027]Alternatively, substituents, R14 and R15 taken together
with the nitrogen atom to which they are bonded form a five-, six-,
seven-, eight-, nine- and ten-membered heterocyclyl moiety.

[0028]In formula I, when Rc is heterocyclyl, then a ring carbon atom
of the heterocyclyl moiety is directly bonded to Z, or in the case where
Z is a bond, to the imidazolyl carbon atom to which Z is bonded.

[0029]The many embodiments of the compounds of formula I of the invention
exhibit useful properties related to their activities as ligands of
cannabinoid receptors and the biological consequences of binding to these
receptors.

[0030]In particular embodiments of the invention, the compounds of formula
I bind one or more cannabinoid receptors, such as without limitation, CB1
and CB2. Such compounds include those that can be classified as agonists,
partial agonists or inverse agonists for a particular cannabinoid
receptor and in certain embodiments these compounds exhibit selectivity
for the CB2 receptor over the CB1 receptor. In one aspect, the
cannabinoid receptor is a mammalian cannabinoid receptor, such as a human
cannabinoid receptor, which can be, but is not limited to, a human CB1 or
CB2 receptor.

[0031]The invention also provides pharmaceutical compositions useful for
the prophylaxis and treatment of a CB2-associated and/or CB1-associated
disease or condition. The pharmaceutical compositions include a compound
of formula I and a pharmaceutically acceptable vehicle, diluent,
excipient or carrier.

[0032]The invention further provides a method of prophylaxis or treatment
of a CB2-associated disease or condition by administering a compound of
formula I or a pharmaceutically acceptable salt, acid salt hydrate,
solvate, stereoisomer, or mixture of stereoisomers thereof. In another
embodiment, the invention provides a method of prophylaxis or treatment
of a CB2-associated and/or CB1-associated disease, disorder or condition
by administering a compound of formula I or a pharmaceutically acceptable
salt, acid salt hydrate, solvate, stereoisomer or mixture of
stereoisomers thereof. Such CB2-associated diseases or conditions and
CB1-associated and CB2-associated diseases, disorders and conditions
include, without limitation, pain and inflammation, wherein such pain can
be inflammatory pain, visceral pain, neuropathic pain or hyperalgesia.
Each of these types of pain can present as acute or chronic pain.

BRIEF DESCRIPTION OF THE FIGURES

[0033]FIG. 1 shows the anti-hyperalgesic effect of intraperitoneal
administration of compound 91 on paw withdrawal threshold (in grams)
after intrapaw administration of Freund's Complete Adjuvant (CFA) as
compared to vehicle alone over a twenty-four hour period after CFA
injection.

[0034]FIG. 2 shows a dose response in the inhibition of acetic
acid-induced writhing in mice, for compounds 317 and 366 administered
subcutaneously at doses of 3 mg/kg, 10 mg/kg and 30 mg/kg.

[0037]The following definitions elucidate the meaning of the listed terms
a used in this specification:

[0038]Alkyl--a saturated branched or straight chain monovalent hydrocarbon
radical of a specified number of carbon atoms. Thus, the term alkyl
includes, but is not limited to, methyl, ethyl, propyl, isopropyl,
n-butyl, t-butyl. A chain of one to six carbon atoms is also herein
interchangeably designated as C1-C6 alkyl; a chain of three to
six carbon atoms can be alternatively designated as C3-C6 alkyl
and so on.

[0039]Alkenyl--refers to branched or straight chain hydrocarbon radical
having at least one double bond between two carbon atoms. It should be
noted that in an alkenyl substituted nitrogen, the unsaturated carbon
atom cannot be bound directly to the nitrogen atom, i.e. there must be at
least one unsaturated carbon (--CH2-- or --CR'R''--) intervening
between the nitrogen atom and the nearest unsaturated carbon atom.

[0040]Alkynyl--refers to branched or straight chain hydrocarbon radical
having at least one triple bond between two carbon atoms. It should be
noted that in an alkynyl substituted nitrogen, the unsaturated carbon
atom cannot be bound directly to the nitrogen atom, i.e. there must be at
least one unsaturated carbon (--CH2-- or --CR'R''--) intervening
between the nitrogen atom and the nearest unsaturated carbon atom.

[0041]Haloalkyl--an alkyl group having one or more hydrogen atoms
substituted with a halogen atom, each independently chosen such that a
haloalkyl group having more than one halogen atom can be a mixed
haloalkyl, such as for instance, 2-fluoro, 2-chloroethyl, or perhalo as
in trifluoromethyl.

[0042]Alkoxy--refers to an (alkyl)a--O-(alkyl)b substituent
group wherein a is zero or an integer, and b is an integer and the alkyl
group is as defined above. So that for instance alkoxy can be and without
limitation, --O-methyl, O-ethyl, --O-propyl, --(CH2)aO-methyl,
--(CH2)aO-ethyl, --(CH2)a--O-propyl, and so forth.

[0043]Cycloalkyl--a saturated monocyclic, polycyclic or bridged
hydrocarbon ring system radical or linking group. In a substituted
cycloalkyl ring, the substituent is bonded to ring carbon atom replacing
a hydrogen atom. The term C3-C10 cycloalkyl is herein used to
designate a ring of three to ten carbon atoms, or a ring of three of more
carbon atoms with the remaining carbon atoms forming one or more alkyl
substituents of the ring. Similarly, a C3-C7 cycloalkyl
designates a saturated or partially unsaturated carbocycle, although not
all the designated number of carbon atoms are necessarily ring carbon
atoms. Cycloalkyl typically includes, but is not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl and
cyclooctyl. However, C10 cycloalkyl includes
1,3,3-trimethylbicyclo[2.2.1]heptyl, wherein seven of the ten designated
carbon atoms form the seven-membered bicyclo-carbocycle and the remaining
three are methyl substituents.

[0044]Cycloalkenyl--partially unsaturated monocyclic, polycyclic or
bridged hydrocarbon ring system radical or linking group having at least
one double bond between two carbon atoms. In a substituted cycloalkenyl
ring, the substituent is bonded to ring carbon atom replacing a hydrogen
atom. The term C3-C10 cycloalkenyl is herein used to designate
a ring of three to ten carbon atoms, or a ring of three or more carbon
atoms with the remaining carbon atoms forming one or more alkyl
substituents of the ring. Similarly, C3-C7 cycloalkenyl
designates as partially unsaturated carbocycle, although not all the
designated number of carbon atoms are necessarily ring carbon atoms.
Cycloalkenyl typically includes, but is not limited to, cyclopentenyl,
cyclohexenyl, cycloheptenyl.

[0045]Heterocyclyl--a saturated, partially unsaturated or unsaturated
monocyclic, polycyclic or bridged hydrocarbon ring system radical or
linking group, wherein at least one ring carbon atom has been replaced
with a heteroatom selected from nitrogen, oxygen and sulfur. A
heterocyclyl moiety system further includes a ring system having one,
two, three or four nitrogen ring atoms, or a ring system having zero,
one, two or three nitrogen ring atoms and one oxygen or sulfur ring atom.
The heterocyclic ring system can include more than one ring heteroatom,
wherein one heteroatom is nitrogen and the other is selected from
nitrogen, oxygen and sulfur. A heterocyclyl moiety is derived by the
removal of one hydrogen atom from a single carbon or nitrogen ring atom.
Heterocyclyl includes, but is not limited to, furyl, thienyl, 2H-pyrrole,
2-pyrrolinyl, 3-pyrrolinyl, pyrrolidinyl, pyrrolyl, 1,3-dioxolanyl,
oxazolyl, thiazolyl, imidazolyl, 2-imidazolinyl, imidazolidinyl,
2-pyrazolinyl, pyrazolidinyl, pyrazolyl, isoxazolyl, isothiazolyl,
oxadiazolyl, triazolyl, thiadiazolyl, tetrazolyl, 2H-pyranyl, 4H-pyranyl,
pyridinyl, piperidinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl,
thiomorpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl,
azepanyl, diazepinyl, indolizinyl, indolyl, isoindolyl, 3H-indolyl,
indolinyl, benzo[b]furyl, benzo[b]thienyl, 1H-indazolyl, benzimidazolyl,
benzothiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl,
cinnolinyl, phthalzinyl, quinazolinyl, quinoxalinyl, 1,8-napthyridinyl,
pteridinyl, quinuclidinyl.

[0046]Heterocyclyl--as used herein, also includes an aromatic heterocycle
such as pyrrolyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, isoxazolyl,
thiazolyl, isothiazolyl, furyl, thienyl, pyridyl, pyrazinyl, pyrimidinyl,
and can be optionally substituted by alkyl. Arylalkyl--an optionally
substituted aryl group attached to the end carbon atom of C1-C4
alkyl group. As used herein "heterocyclyl" also includes bicyclic
heterocyclyl moieties in which one or both rings are heterocyclic, such
as for example, but not limited to imidazopyrazinyl, benzofuranyl,
benzodioxolyl, benzothiophenyl, and quinolinyl.

[0047]Aryl--an unsaturated, π-electron conjugated monocyclic or
polycyclic hydrocarbon ring system radical or linking group of six,
eight, ten or fourteen carbon atoms. An aryl radical is derived by the
removal of one hydrogen atom from a single carbon ring atom. Aryl
includes, but is not limited to, phenyl, naphthalenyl, azulenyl,
anthracenyl.

[0048]Aminosulfonylalkyl--a radical of the formula --NHSO2-alkyl.
Sulfonyl-aminoalkyl--a linking group of the formula --SO2NH-alkyl-
or a radical of the formula --SO2N(alkyl)2. Alkylcarbamoyl--a
linking group of the formula -alkyl-C(O)NH-- or a radical of the formula
-alkyl-C(O)NH2. Carbamoylalkyl--a linking group of the formula
--NHC(O)-alkyl- or a radical of the formula --NHC(O)-alkyl.
Halogen--fluoro, chloro, bromo or iodo. Carboxyl--a radical of the
formula --COOH. Hydroxyl--a radical of the formula --OH. Cyano--a radical
of the formula --C≡N. Oxo--a radical of the formula ═O in which
the oxygen atom is double bonded. Amino--a radical of the formula
--NH2 or a linking group having the formula --NH--. Aminoalkyl--a
radical of the formula --NH-alkyl or --N(alkyl)2.

[0049]As used herein, the terms: compound, salt, polymorph, isomer,
solvate are also interchangeably referred to in the plural form (i.e.
compounds, salts, polymorphs, isomers and solvates).

[0050]The compounds of the present invention can contain one or more
stereogenic centers, depending upon the location and nature of the
various substituents desired. These stereogenic centers may be present in
the (R) or (S) configuration, resulting in racemic mixtures and/or
diastereomeric mixtures. Substituents on a partially or fully saturated
ring may also be present in either cis or trans form. All such
configurations (including enantiomers and diastereomers) of the compounds
described or exemplified herein, are contemplated within the scope of the
present invention. Compounds of the invention can also exist as
individual stereoisomers or as mixtures in varying ratios (e.g.
enantiomerically enriched or racemates). Enantiomeric mixtures of the
compounds may be partially or fully resolved through standard
purification and/or separation techniques known in the art, including but
not limited to chiral chromatography (e.g. chiral derivatized solid
phase), formation and separation of diastereomeric salts (e.g. tartaric
acid salts or camphorsulfonic acid salts), or enzymatic separation.
Diastereomeric mixtures can be separated by techniques well known in the
art, based on their physical and/or chemical differences, or by methods
described above.

[0051]In this specification, salts of a compound of formula I refers to a
complex of the compound with an inorganic or organic counter ion or
counter ions. For examples, see Handbook of Pharmaceutical Salts:
Properties, Selection and Use; Stahl P. H., Wermuth, C. G., Eds.; John
Wiley and Sons, 2002. Pharmaceutically useful salts include those
obtained by treating the compound, functioning as a base, with an
inorganic or organic acid to form a salt or salts. Additional
pharmaceutically useful salts include those obtained by treating the
compound, functioning as an acid, with an inorganic or organic base to
form a salt or salts. Other pharmaceutically useful salts include those
obtained by treatment of basic nitrogen-containing groups with such
agents as alkyl halides such as chlorides or bromides to form a
quaternary ammonium a salt or salts.

[0052]As used herein, the term "solvates" describes a complex wherein the
compound is coordinated with a proportional amount of a solvent molecule.
Specific solvates, wherein the solvent is water, is referred to as
hydrates. Combinations of a drug and propylene glycol (1,2-propanediol)
have been used to form pharmaceutical drug solvates. See for example U.S.
Pat. No. 3,970,651. Other suitable solvates are hydrates of drug
compounds. Such hydrates include hydrates which either have comparable
activity or hydrates which are converted back to the active compound
following administration.

[0053]The compounds of the present invention described and exemplified
herein modulate a signal that regulates a biological activity, by
modulating the activity of a cannabinoid receptor. Modulation of a
cannabinoid receptor can be effected by a compound of the present
invention acting as an agonist, a partial agonist, inverse agonist or an
antagonist upon binding at a cannabinoid receptor such as CB2 and/or CB1.
The modulation of a cannabinoid receptor can be activation by compound of
the present invention acting an agonist. Alternatively, the modulation of
a cannabinoid receptor can be inhibition or deactivation by an
antagonist. One particular signal regulated by CB2 is the intracellular
concentration of cyclic adenosine monophosphate (cAMP).

[0054]The term `agonist` as used herein means a molecule that produces a
physiological response by activating a receptor. The term `inverse
agonist` as used herein means a molecule that tends to reverse the effect
of an agonist. Current theory holds that this occurs due to the higher
affinity of the inverse agonist for binding the inactive conformation
over the active conformation of the receptor. The term `antagonist` as
used herein means a molecule that binds a receptor and thereby interferes
with the interaction of an agonist and its cognate receptor, or blocks
the constitutive activity of the receptor. The term `neutral antagonist`
as used herein means a molecule that binds a receptor with equal affinity
for the active and inactive conformations and thereby inhibits receptor
activity by competing with an agonist.

[0055]The compounds useful in the practice of the present invention have
the structure of formula I:

##STR00005##

[0056]In particular embodiments of the invention, Y is an amino-radical,
NRa or a quaternary amino radical N+R1R2 with an
anionic counterion X.sup.-. The anionic counterion X.sup.- can be any
anionic counterion, such as for instance, an inorganic counterion such as
chloride, or an organic counterion such as succinate; and m is an integer
equal to 1, 2 or 3, such that the Y-containing ring includes six, seven
or eight ring atoms fused to the imidazole ring. Z is a bond or a
bivalent linking group chosen from --(CH2)p--, --CH═CH--,
--C≡C--, --CONH-- and --CO--; wherein p is an integer from 1 to 6.

[0057]The compounds have the structure of formula I, wherein Ra is
hydrogen or a substituent chosen from C1-C8 alkyl,
C3-C6 alkenyl, C3-C6 alkynyl, aryl, C3-C8
cycloalkyl, C3-C8 cycloalkenyl, --SO2R3, --COR3,
--CONR3R4, --CSNR3R4, --COOR3, and
--(CH2)q-linked-heterocyclyl; wherein q is zero or an integer
from one to four. The alkyl, cycloalkyl, cycloalkenyl, aryl and
heterocyclyl substituents of Ra are optionally substituted with from
one to four groups, each independently chosen from halo, hydroxyl, oxo,
amino, nitro, cyano, carboxyl, --COR3, C1-C6 alkyl,
C1-C4 alkoxy, C3-C8 cycloalkyl, phenyl,
trifluoromethoxy and trifluoromethyl. In one embodiment when Ra is
--SO2R3, then R3 is not hydrogen.

[0058]In the compounds having the structure of formula I, Rb is a
radical bonded through the carbonyl to the imidazolyl ring. Rb is
chosen from C1-C8 alkyl, C2-C8 alkenyl, aryl,
--NR5R6, 4-R7-substituted piperazinyl, and
4-R8,4-R9-substituted piperidinyl; wherein the alkyl, alkenyl
and aryl are optionally substituted with from one to three groups chosen
independently from the following: C1-C4 alkyl, C2-C4
alkenyl, C2-C6 alkynyl, C3-C10 cycloalkyl,
C1-C4 alkoxy, aryl, five-membered, six-membered and
seven-membered heterocyclyl, halo, hydroxyl, amino, cyano and nitro.

[0060]In formula I, when Rc is heterocyclyl, the heterocyclyl moiety
is directly bonded through a carbon atom of the heterocyclic ring or ring
system to the radical Z, or if Z is a bond, to the imidazole ring of
formula I.

[0061]The substituents, R1 and R2 are each C1-C4
alkyl. The substituents, R1 and R2 can be identical or
different, branched or straight chain alkyl substituents.

[0062]The substituents, R3 and R4 are each independently chosen
from the following: hydrogen, C1-C6 alkyl, C3-C8
alkenyl, C3-C8 alkynyl, C3-C8 cycloalkyl,
C3-C8 cycloalkenyl, aryl, and heterocyclyl having from four to
eight ring atoms. Each R3 and R4 can be optionally substituted
with one to three groups independently chosen from C1-C6 alkyl,
C1-C6 haloalkyl, C3-C8 cycloalkyl, C1-C4
alkoxy, C1-C4 acyl, aryl, five- to eight-membered monocyclic
heterocyclyl, 9-, 10-membered bicyclic heterocyclyl, amino, nitro, cyano,
hydroxyl, carboxyl, oxo, and halo. However, when Ra is
--SO2R3, then R3 is not H.

[0063]Alternatively, R3 and R4 can be taken together with the
nitrogen atom to which they are bonded to form a heterocyclyl moiety,
wherein the heterocyclyl formed from R3 and R4 can be a
four-membered heterocyclyl, a five-membered heterocyclyl, a six-membered
heterocyclyl, a seven-membered heterocyclyl or an eight-membered
heterocyclyl moiety.

[0064]The substituent, R5 is hydrogen or a substituent chosen from
the following: C1-C4 alkyl, and C1-C4 haloalkyl. The
alkyl and haloalkyl of R5 are optionally substituted with one to
four substituents independently chosen from C1-C4 alkoxy,
hydroxyl, amino and cyano.

[0065]The substituent, R6 is hydrogen or a substituent chosen from
the following: --CR10R11R12,
--CR10R11COR13, C1-C8 alkyl, C3-C10
cycloalkyl, aryl, and heterocyclyl; wherein the alkyl, cycloalkyl, aryl,
and 5-, 6-, 7-, 8-membered monocyclic heterocyclyl, and 9-, 10-membered
bicyclic heterocyclyl can be optionally substituted by from one to three
substituents independently selected from the group consisting of
C1-C4 alkyl, aryl, halo, --OH, C1-C4 alkoxy,
--NH2, --CN, --NO2.

[0066]Alternatively, the substituents, R5 and R6 taken together
with the nitrogen atom to which they are bonded form a 5-, 6-, 7-,
8-membered monocyclic heterocyclyl, and 9-, 10-membered bicyclic
heterocyclyl, which monocyclic heterocyclyl, or bicyclic heterocyclyl is
optionally substituted with one or two substituents independently chosen
from oxo and --CONR1R2.

[0069]In a first alternative, the substituents, R8 and R9 taken
together with the nitrogen atom to which they are bonded form a
heterocyclyl moiety, which heterocyclyl moiety is optionally substituted
with one to three substituents chosen from C1-C4 alkyl, halo,
oxo and aryl.

[0070]In a second alternative, the substituents, R8 and R9 taken
together with the carbon atom to which they are bonded form a carbocyclyl
ring, which heterocyclyl moiety is optionally substituted with 1-3
substituents chosen from C1-C4 alkyl, halo, oxo and aryl.

[0072]The substituent, R12 is chosen from hydrogen, C1-C4
alkyl and C1-C4 hydroxyalkyl; and the substituent, R13 is
chosen from --OR10 and --NR8R9.

[0073]The substituents, R14, R15 and R16 are each
independently hydrogen or C1-C4 alkyl; or alternatively,
substituents, R14 and R15 taken together with the nitrogen atom
to which they are bonded form a five-membered, six-membered,
seven-membered and eight-membered monocyclic heterocyclyl, nine-membered
and ten-membered bicyclic heterocyclyl moiety.

[0074]In one embodiment of the invention, in the compounds of formula I, Y
is NRa or N+R1R2 X.sup.-, wherein X.sup.- is a halide
ion; and Ra is chosen from the following: hydrogen, C1-C6
alkyl, cyclopropyl, --SO2R3, --COR3, --CONR3R4,
--CSNR3R4, --CO2R3, and
--(CH2)pheterocyclyl, wherein p is zero or 1; and m is 1 or 2;
and the alkyl, aryl and heterocyclyl of Ra are each optionally
substituted with halo, hydroxyl, cyclopropyl, acetyl or phenyl. In this
embodiment, the substituent, R3 is chosen from C1-C5
alkyl, cyclopropyl, five-membered heterocyclyl, six-membered heterocyclyl
and aryl; wherein the aryl substituent of Ra is optionally
substituted with cyano, nitro, halo or trifluoromethyl.

[0075]In one particular aspect of this embodiment, the radical Ra is
hydrogen, C1-C4 alkyl, 4-fluorophenyl-sulfonyl, or
--(CH2)p-pyrimidinyl, wherein the alkyl of Ra, is
optionally substituted with cyclopropyl.

[0076]In another embodiment of the compounds of formula I, the radical
Rb is chosen from C1-C6 alkyl, C7-C6 alkenyl,
NR5R6,

##STR00006##

wherein the alkyl of Rb is optionally substituted with aryl and
R3 is aryl and R5 is hydrogen. The substituent, R6 is
chosen from the following: --CR10R11R12,
--CR10R11COR13, C1-C6 alkyl, C3-C10
cycloalkyl, aryl, and five-membered, six-membered, seven-membered and
eight-membered monocyclic heterocyclyl, nine-membered and ten-membered
bicyclic heterocyclyl. The alkyl, cycloalkyl, aryl, and heterocyclyl
substituents of R6 are themselves optionally substituted with from
one to three substituents independently chosen from: methyl, aryl, halo,
and hydroxyl. Additionally, in this embodiment, the heterocyclyl of
R6 is optionally substituted with a single --CONHR1R2
substituent. The substituent, R7 is either --COR3 or a
six-membered heterocyclyl. Substituents, R8 and R9 are
independently chosen from: hydrogen, C1-C4 alkyl,
C1-C2 haloalkyl, C1-C3 alkoxyalkyl, C3-C4
cycloalkyl, --CONH2, five-membered monocyclic heterocyclyl,
six-membered monocyclic heterocyclyl and nine-membered bicyclic
heterocyclyl, and ten-membered bicyclic heterocyclyl; wherein the
C1-C4 alkyl and five-, and six-membered monocyclic heterocyclyl
of R8 and R9 are optionally substituted with a six-membered
monocyclic heterocyclyl, or one or two methyl groups. Alternatively,
R8 and R9, taken together with the atom to which they are
bonded form a carbocyclic or heterocyclyl moiety, which carbocyclic or
heterocyclyl moiety is optionally substituted with one to two
substituents independently chosen from methyl, halo, oxo and aryl. The
substituent, R10 in this embodiment is either hydrogen or
C1-C4 alkyl; and the substituent, R11 is chosen from:
hydrogen, C1-C5 alkyl, C3-C10 cycloalkyl, aryl,
C1-C4 alkylaryl, and five-membered and six-membered monocyclic
heterocyclyl; wherein the alkyl, cycloalkyl, aryl, and heterocyclyl of
R11 are optionally substituted with from one to three substituents
independently chosen from C1-C4 alkyl, C3-C6
cycloalkyl, aryl, five-membered heterocyclyl, 6-membered heterocyclyl,
and nine-membered bicyclic heterocyclyl, halo, hydroxyl, --COOR10,
--CONR8R9, and --SO2NR8R9.

[0077]In one aspect of this embodiment, the radical, Rb is
NR5CHR11COR13. In a particular example of the aspect
wherein Rb is NR5CHR11COR13, the substituent, R5
is hydrogen and R13 is NR8R9. In another example of this
aspect, the substituent, R8 is hydrogen and R9 is methyl. In a
particular aspect of this embodiment, the radical, Rb is
--NHCH(tBu)CONHCH3.

[0079]In another embodiment of the compounds of formula I, Z is a bond, or
Z is --(CH2)p, or --CH═CH--; and the radical Rc is
chosen from C3-C8 cycloalkyl, C3-C8 cycloalkenyl,
phenyl, five-membered heterocyclyl and six-membered heterocyclyl, wherein
the cycloalkyl, cycloalkenyl, phenyl and heterocyclyl of Rc are
optionally substituted with from one or two substituents independently
chosen from C1-C4 alkyl, C1-C4 alkoxy,
C3-C6 cycloalkyl, halo, trifluoromethoxy, trifluoromethyl,
hydroxyl, cyano, and an additional optional independently selected halo
substituent.

[0080]In one aspect of the above embodiment of the compounds of formula I,
Z is a bond and the radical Rc is optionally substituted phenyl,
wherein the phenyl of Rc is optionally substituted with from one or
two substituents independently chosen from halo, methyl, methoxy,
trifluoromethyl and cyano; and the phenyl of Rc is further
optionally substituted with an additional halo substituent. In a
particular aspect of this embodiment, the radical Rc is one of the
following: phenyl, 3-chloro-4-methylphenyl, 2-chloro-4-fluorophenyl,
2-fluoro-4-chlorophenyl, 2-fluoro-4-bromophenyl, 2-fluoro-5-chlorophenyl,
2,4-difluorophenyl, 2,5-difluorophenyl, 3,5-difluorophenyl,
3,4-difluorophenyl, 2-fluoro-4-methylphenyl, 2-fluoro-5-methylphenyl,
2-fluoro-3-methoxyphenyl, 2-fluoro-4-methoxyphenyl,
2-fluoro-4-trifluoromethylphenyl, 2-fluoro-5-trifluoromethylphenyl,
3-cyano-4-fluorophenyl, 2-fluoro-4-methyl-5-chlorophenyl,
2,4-difluoro-5-chlorophenyl, 2,4,5-trifluorophenyl,
3,4,5-tri-fluorophenyl, 2,5-difluoro-4-methoxyphenyl, 2-fluorophenyl,
3-fluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3-chlorophenyl,
4-chlorophenyl, 3-methyl-4-fluorophenyl, 2-fluoro-3-chlorophenyl,
3-trifluoromethyl-phenyl, 3-methylphenyl, 3-fluoro-4-methylphenyl,
3-methyl-4-fluorophenyl, 3-chloro-4-fluorophenyl and
3-fluoro-4-chlorophenyl.

[0081]In a particular aspect of the above embodiment of the compounds of
formula I, Z is a bond and the radical Rc is chosen from phenyl,
2-fluoro-4-chlorophenyl, 2-fluoro-4-bromophenyl,
2,4-fluoro-5-chlorophenyl, and 2,4,5-trifluorophenyl.

[0082]In another aspect of the above embodiment of the compounds of
formula I, Z is a bond and the radical Rc is chosen from the
following: C2-C6 alkyl, C2-C8 alkenyl,
C3-C8 cycloalkyl, C3-C8 cycloalkenyl, five-membered
heterocyclyl, and six-membered heterocyclyl. In this aspect of the above
embodiment of the compounds of formula I, the alkyl, cycloalkyl,
cycloalkenyl and heterocyclyl of Rc are optionally substituted with
from one to two substituents independently chosen from C1-C4
alkyl, methoxy, trifluoromethyl, C3-C6 cycloalkyl, halo,
hydroxyl and cyano.

[0085]The present invention further provides pharmaceutically acceptable
salts, acids salts, solvates (including hydrates) and stereoisomers of
the compounds having the structure of formula I. Also provided are
mixtures of stereoisomers of the compounds having the structure of
formula I wherein the mixture can include equal quantities of each
stereoisomer, or the mixture can contain an excess of one stereoisomer
over another.

[0086]In one embodiment of the invention, the compounds having the
structure of formula I bind one or more cannabinoid receptors such as,
without limitation the CB1 or CB2 receptor. Certain compounds of the
invention exhibit an EC50 for the CB1 receptor and/or the CB2
receptor of from about 0.1 nM to about 10 μM; or from about 1 nM to
about 1 μM; or from about 5 nM to about 500 nM.

[0087]As used herein, a cannabinoid receptor-associated disease, disorder
or condition is any disease, disorder or condition that is preventable or
treatable by modulation of a cannabinoid receptor, such as and without
limitation, CB2 or CB1. The modulation can be activation by an agonist,
or inhibition by an inverse agonist. The cannabinoid receptor can be any
mammalian cannabinoid receptor, such as but not limited to, a human
cannabinoid receptor or a rat cannabinoid receptor. In one aspect, the
compounds of the invention having the structure of formula I are
cannabinoid receptor agonists that activate a cannabinoid receptor.

[0088]The cannabinoid receptor-associated disease, disorder or condition
can be any cannabinoid receptor-associated disease, disorder or
condition, such as and without limitation: pain, inflammation,
immunomodulation and pruritis; and can also include osteoclastogenesis.
The cannabinoid receptor-associated disease, disorder or condition can
also be obesity.

[0090]The cannabinoid receptor-associated inflammation can be otic or
ocular inflammation due to any of a variety of causes; inflammation due
to rheumatoid arthritis, eczema, atopic dermatitis, inflammatory bowel
disease, irritable bowel syndrome, kidney dialysis, insect bites or the
inflammation can be inflammation caused by autoimmunity.

[0091]The cannabinoid receptor-associated pruritis can be opioid-induced
pruritis, where in the pruritis is caused by use or abuse of an opioid,
such as morphine.

[0092]The cannabinoid receptor can be any mammalian cannabinoid receptor,
such as but not limited to, a human cannabinoid receptor or a rat
cannabinoid receptor. In one aspect, the compounds of the invention
having the structure of formula I are cannabinoid receptor agonists that
activate a cannabinoid receptor.

[0093]In some embodiments, a particular dose and route of administration
of the compound can be chosen by a clinician to completely prevent or
cure the disease, disorder or condition. In other embodiments a
particular dose and route of administration of the compound chosen by the
clinician ameliorates or reduces one or more symptoms of the disease,
disorder or condition.

[0094]As used herein, "effective amount" or "sufficient amount" of the
compound of the invention refers to an amount of the compound as
described herein that may be therapeutically effective to inhibit,
prevent, or treat a symptom of a particular disease, disorder, condition,
or side effect.

[0095]As used herein, "pharmaceutically acceptable" refers to compounds,
materials, compositions, and/or dosage forms which are, within the scope
of sound medical judgment, suitable for contact with the tissues of human
beings and animals without severe toxicity, irritation, allergic
response, or other complications, commensurate with a benefit-to-risk
ratio that is reasonable for the medical condition being treated.

[0096]As used herein, a "pharmaceutically acceptable salt" refers to a
derivative of a compound wherein the parent compound is modified by
making an acid or a base salt thereof. Examples of pharmaceutically
acceptable salts include, but are not limited to, mineral or organic acid
salts of basic residues such as amines; alkali or organic salts of acidic
residues such as carboxylic acids and the like.

[0097]The pharmaceutically acceptable salts include the conventional
non-toxic salts or the quaternary ammonium salts of the parent compound
formed, for example, from non-toxic inorganic or organic acids. For
instance, such conventional non-toxic salts include those derived from
inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic,
phosphoric, nitric acids and the like; and the salts prepared from
organic acids such as acetic, propionic, succinic, glycolic, stearic,
lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic,
phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic,
fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic,
isethionic acids, and the like. These physiologically acceptable salts
are prepared by methods known in the art, e.g., by dissolving the free
amine bases with an excess of the acid in aqueous alcohol, or
neutralizing a free carboxylic acid with an alkali metal base such as a
hydroxide, or with an amine. Thus, a pharmaceutically acceptable salt of
a substituted imidazoheterocycle of the invention can be formed from any
such compound having either acidic, basic or both functional groups. For
example, a compound having a carboxylic acid group, may in the presence
of a pharmaceutically suitable base, form a carboxylate anion paired with
a cation such as a sodium or potassium cation. Similarly, a compound
having an amine functional group may, in the presence of a
pharmaceutically suitable acid such as HCl, form a salt.

[0100]Buffers include phosphate and citrate. Antioxidants include sodium
bisulfite. Local anesthetics include procaine hydrochloride. Suspending
and dispersing agents include sodium carboxymethylcelluose, hydroxypropyl
methylcellulose and polyvinylpyrrolidone. Emulsifying agents include
Polysorbate 80 (Tween 80). A sequestering or chelating agent of metal
ions such as EDTA can also be incorporated. Pharmaceutical carriers also
include ethyl alcohol, polyethylene glycol and propylene glycol for water
miscible vehicles; the pH can be adjusted to a physiologically compatible
pH by addition of sodium hydroxide, hydrochloric acid, citric acid or
lactic acid.

[0101]The pharmaceutical compositions that include the compounds of
formula I of the invention can be delivered or administered
intravenously, transdermally, transmucosally, intranasally,
subcutaneously, intramuscularly, orally or topically (such as for example
to the eye). The compositions can be administered for prophylaxis or
treatment of individuals suffering from, or at risk of a disease,
disorder or condition. Prophylaxis is defined as a measure designed to
preserve the health of an individual.

[0102]For therapeutic applications, a pharmaceutical composition is
typically administered to a subject suffering from a disease, disorder or
condition, in an amount sufficient to inhibit, prevent, or ameliorate the
disease, disorder or condition. An amount adequate to accomplish this is
defined as a "therapeutically effective dose."

[0103]The pharmaceutical compositions of the invention can be administered
to a mammal for prophylactic or therapeutic purposes in any of the
above-described formulations and delivery modes. The mammal can be any
mammal, such as a domesticated or feral mammal, or even a wild mammal.
The mammal can be any mammal, such as for instance a primate, ungulate,
canine or feline. For instance, and without limitation, the mammal can be
a pet or companion animal, such as a dog or a cat; a high-value mammal
such as a thoroughbred or show animal; a farm animal, such as a cow, a
goat, a sheep or pig; or a primate such as an ape or monkey. In one
embodiment, the mammalian cannabinoid receptor is a human cannabinoid
receptor, such as a human CB1 or a human CB2 receptor.

[0104]Without wishing to be bound by any particular theory, it is believed
that due to their ability to bind and modulate the activity of the CB1
receptor and/or the CB2 receptor, the compounds of the present invention
are useful in the treatment of diseases, disorders or conditions that
include, but are not limited to, inflammatory diseases such as rheumatoid
arthritis, systemic lupus erythematosus, Crohn's disease, psoriasis,
eczema, multiple sclerosis, diabetes and thyroiditis.

[0106]In addition, certain compounds of the invention can be used to
modulate bone formation and/or resorption for treating conditions
including, but not limited to, ankylosing spondylitis, gout, arthritis
associated with gout, osteoarthritis and osteoporosis. Certain compounds
of the invention can also be used for the treatment of neuropathic pain
including but not limited to diabetic neuropathy, fibromyalgia, lower
back pain, sciatica, pain from physical trauma, cancer, amputation,
toxins or chronic inflammatory conditions.

[0107]The compounds of the invention and their pharmaceutically acceptable
salts can be administered in a standard manner, for example orally,
parentarally, sublingually, dermally, transdermally, rectally, via
inhalation, or by buccal, nasal, ocular or otic administration.

General Methods

[0108]All reactions involving moisture sensitive compounds were carried
out under an anhydrous nitrogen or argon atmosphere. All reagents were
purchased from commercial sources and used without further purification.
Unless otherwise noted, the starting materials used in the examples were
obtained from readily available commercial sources or synthesized by
standard methods known to those skilled in the art of organic synthesis.
Reactions performed under microwave irradiation conditions were carried
out in a Biotage Initiator® 60 microwave system (Charlottesville,
Va.; model no. 10986-22V) with a 300 Watt magnetron. Normal phase
chromatography and reverse phase chromatography was performed on an ISCO
CombiFlash® Companion®, CombiFlash® Companion/TS® system
(Teledyne Isco, Inc., Lincoln, Nebr.) or ISCO CombiFlash® Sq
16×. Reverse phase chromatography was also performed on a Waters
Autopurification System with 3100 Mass Detector. The HPLC column was a
Waters XBridge C18 5 μm OBD 19×150 mm; eluents were A: water
with 0.1% formic acid and B: acetonitrile with 0.1% formic acid. Gradient
elution was from 5% B-95% B. The total run time was 13 mins. Mass spectra
(MS) data were acquired on the Waters SQ Detector/3100 Mass detector
using electrospray techniques or a Waters ZQ mass spectrometer with a
Waters 600 HPLC pump and a 2487 UV detector and a 1525u binary LC pump
with integrated degasser.

[0109]Compounds were also characterized by their
LCMS-Electrospray/chemical ionization mass spectra (LC ESCI-MS) on one of
the following systems:

[0110](1) Waters HPLC-MS system (Waters Corp., Milford, Mass.) equipped
with a 2767 Sample Manager, 2545 Binary Gradient Module, SFO System
Fluidics Organizer, 2996 Photodiode Array Detector and 3100 Mass
Detector. Data were collected across a range of wavelengths from 220-280
nm in positive ESCI mode. Spectra were scanned from 100-1400 atomic mass
units (amu). The HPLC column was a Waters XBridge C18 3.5 μm
4.6×30 mm; eluents were A: water with 0.1% formic acid and B:
acetonitrile with 0.1% formic acid. Gradient elution was from 5% B-95% B
over 2.3 minutes with an initial hold of 0.2 minutes and a final hold at
95% B of 0.5 minutes. The total run time was four minutes.

[0111](2) Waters (Waters Corporation, Milford, Mass.) UPLC-MS system
equipped with an Acquity Sample Manager, Acquity Binary Solvent Manager,
Acquity Photodiode Array Detector, Acquity Evaporative Light Scattering
Detector and SQ Detector. Data were collected at 220 nm and 254 nm and in
positive electrospray-chemical ionization mode. The UPLC column used was
a Waters Acquity UPLC BEH C18 1.7 um 2.1×50 mm. Spectra were
scanned from 100-1400 amu. The eluents were A: water with 0.1% formic
acid and B: acetonitrile with 0.1% formic acid. Gradient elution from 5%
B to 95% B over 0.8 minutes was used with a final hold at 95% B of 0.2
minutes at a flow rate of 0.8 milliliters per minute. Total run time was
1.5 minutes.

[0114]General schemes for the preparation of intermediates used in the
synthesis of the compounds of the invention detailed below are described
in detail in U.S. Pat. No. 7,517,874 which is hereby incorporated by
reference.

[0116]Step 1: Preparation of methyl
4-(diethoxymethyl)-1H-imidazole-5-carboxylate (Intermediate 1B). To a
stirred suspension of 30-35% KH (7.90 g) in 40 mL anhydrous diglyme at
-20° C. was added a solution of diethoxyacetonitrile (Intermediate
1A, 6.20 g, 46.6 mmol) and methyl isocyanoacetate (4.96 g, 65.2 mmol) in
25 mL of anhydrous diglyme. The resulting mixture was heated to
70-80° C. and stirred overnight. The mixture was cooled to room
temperature and quenched with saturated NH4Cl solution. DCM was
added and the layers were separated. The mixture was further extracted
with DCM. The combined organic extracts were dried over anhydrous
MgSO4, filtered and concentrated under reduced pressure to give
brown oil. Cold ether was added to the residue and the resulting white
precipitate was filtered, washed with cold ether and dried to give the
desired product Intermediate 1B (5.65 g, 53%) as a white solid.

[0117]Step 2: Preparation of methyl 4-formyl-1H-imidazole-5-carboxylate
(Intermediate 1C). To a solution of Intermediate 1B (5.65 g, 24.75 mmol)
in water (12 mL) was added acetic acid (49 mL, 0.86 mol). The resulting
mixture was stirred under nitrogen for 6 hours. The reaction mixture was
azeotroped with toluene and dried under vacuum to give the desired
aldehyde, Intermediate 1C in quantitative yield as a white solid, which
was used in the next step without further purification.

[0118]Step 3: Preparation of methyl
4-((benzyl(2-hydroxyethyl)amino)methyl)-1H-imidazole-5-carboxylate
(Intermediate 1D). To a stirred suspension of Intermediate 1C (3.20 g,
20.76 mmol) in dry THF (180 mL) was added anhydrous Na2SO4
(14.48 g, 192 mmol) and N-benzylethanolamine (3.70 g, 24.47 mmol). The
resulting mixture was stirred at room temperature under nitrogen for 1
hour. Sodium triacetoxyborohydride (6.37 g, 28.5 mmol) was added
portion-wise and the resulting mixture was stirred under nitrogen for 48
hours. The resulting mixture was quenched with water and neutralized with
saturated NaHCO3 solution. The mixture was extracted with DCM and
the combined organic extracts were dried over anhydrous Na2SO4,
filtered and concentrated. The crude mixture was purified using normal
phase chromatography eluting with a 10-30% methanol/DCM gradient to
provide Intermediate 1D as a white solid (5.80 g, 98%).

[0119]Step 4: Preparation of methyl
4-((benzyl(2-chloroethyl)amino)methyl)-1H-imidazole-5-carboxylate
hydrochloride (Intermediate 1E). To a solution of Intermediate 1D (0.94
g, 3.24 mmol) in DCM (30 mL) was added thionyl chloride (0.95 mL, 12.96
mmol). The resulting mixture was stirred at 44° C. overnight and
allowed to cool to ambient temperature. The mixture was concentrated
under reduced pressure, azeotroped with acetonitrile and dried under
vacuum overnight to give Intermediate 1E in quantitative yield as a white
solid which was used in the next step without further purification.

[0120]Step 5: Preparation of methyl
7-benzyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate
(Intermediate 1F). The chloride, Intermediate 1E (0.97 g, 3.15 mmol) was
dissolved in acetonitrile (30 mL) and TEA (1.77 mL, 12.62 mmol) was added
drop wise. The resulting mixture was stirred at 80° C. under
nitrogen overnight. The mixture was allowed to cool, was filtered and the
filtrate was concentrated. The residue was partitioned between DCM and
saturated NaHCO3 solution and the phases were separated. The aqueous
phase was further extracted with DCM and combined organic extracts were
dried over anhydrous MgSO4, filtered and concentrated. The crude
residue was purified using normal phase chromatography, eluting with a
0-40% methanol/DCM gradient to provide product Intermediate 1F (0.58 g,
66%) as a brown solid.

[0121]Step 6: Preparation of 7-tert-butyl 1-methyl
5,6-dihydroimidazo[1,5-a]pyrazine-1,7(8H)-dicarboxylate (Intermediate
1G). Under a nitrogen atmosphere, the product Intermediate 1F (3.70 g,
13.64 mmol) was dissolved in ethanol (180 mL) and
di-tert-butyldicarbonate (3.87 g, 17.73 mmol) was added followed by DIEA
(7.15 mL, 40.9 mmol) and 20% palladium hydroxide on carbon (1.92 g, 2.73
mmol). The resulting black suspension was stirred under a hydrogen
atmosphere (90 psi) for 48 hours using a Parr hydrogenator. The mixture
was filtered through a pad of celite and washed with methanol. The
filtrate was concentrated, dissolved in ethyl acetate and washed with
saturated NaHCO3 solution and brine. The organic layer was dried
over anhydrous Na2SO4, filtered and concentrated to give the
desired product Intermediate 1G as a white solid (3.20 g, 83%) which was
used in the next step without further purifications.

[0122]Step 7: Preparation of 7-tert-butyl 1-methyl
3-bromo-5,6-dihydroimidazo[1,5-a]pyrazine-1,7(8H)-dicarboxylate
(Intermediate 1H). The Intermediate 1G (3.20 g, 11.38 mmol) was dissolved
in acetonitrile and NBS (2.43 g, 13.65 mmol) was added. The reaction
mixture was stirred at room temperature overnight. The mixture was
concentrated and partitioned between ethyl acetate and water. The organic
layer was washed with brine, dried over anhydrous Na2SO4,
filtered and concentrated to give a yellow solid. The solid was dissolved
in DCM and passed through a silica gel plug, eluting with 10% methanol in
DCM to give product Intermediate 1H as a yellow solid (3.90 g, 95%).

[0123]Step 8: Preparation of
3-bromo-7-(tert-butoxycarbonyl)-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-
-1-carboxylic acid (Intermediate 1I). The product Intermediate 1H (0.85 g,
2.36 mmol) was dissolved in methanol (50 mL) and LiOH (0.79 g, 18.88
mmol) in water (10 mL) was added. The resulting solution was stirred at
50° C. overnight. The reaction mixture was concentrated, cooled on
ice and brought to pH 3 using 1N HCl. The resulting white precipitate was
filtered, washed with water and air dried to give the desired acid
Intermediate 1I as a white solid (0.62 g, 76%).

[0124]Step 9: Preparation of (S)-tert-butyl
3-bromo-1-(3,3-dimethyl-1-(methylamino)-1-oxobutan-2-ylcarbamoyl)-5,6-dih-
ydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate (Intermediate 1J). Acid
Intermediate 1I (0.62 g, 1.79 mmol) was dissolved in DMF and
L-tert-Leucine methylamide (0.31 g, 2.14 mmol) was added followed by DIEA
(0.94 mL, 5.37 mmol). The resulting mixture was stirred for 20 minutes,
HBTU (0.75 g, 1.97 mmol) was added in one portion and the mixture was
stirred overnight. The mixture was diluted with water and extracted with
ethyl acetate. The organic layer was washed successively with water, then
brine, and dried over anhydrous Na2SO4, filtered and
concentrated. The residue was purified by normal phase chromatography,
eluting with a 0-10% methanol/DCM gradient to provide product
Intermediate 1J as an off-white solid (0.68 g, 80%).

[0125]Step 10: Preparation of (S)-tert-butyl
1-(3,3-dimethyl-1-(methylamino)-1-oxobutan-2-ylcarbamoyl)-3-phenyl-5,6-di-
hydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate (Compound 1). Intermediate
1J (0.20 g, 0.42 mmol) was dissolved in dioxane (4 mL) and phenylboronic
acid (0.10 g, 0.85 mmol) was added, followed by 2N Na2CO3
solution (0.70 mL, 1.39 mmol). The resulting mixture was degassed with
nitrogen and tetrakis(triphenylphosphine) palladium(0) (0.073 g, 0.06
mmol) was added. The mixture was heated in a microwave reactor at
150° C. for 20 min. The reaction mixture was filtered through a
celite pad, rinsed with methanol and the combined filtrate and washings
were concentrated. The residue was purified by normal phase
chromatography, eluting with 0-100% hexanes/ethyl acetate to provide
Compound 1 as a yellow solid (0.15 g, 75%).

[0127]Additional Compounds 3-21 were synthesized by the same procedure as
described above except that alternative boronic acids were used in place
of phenylboronic acid in the reaction with Intermediate 1J to form
Intermediate 1. For example, Compound 18 was synthesized using
3-chlorophenyl boronic acid. These intermediates were then deprotected
with TFA as described in Example 2 to form additional Compounds 3-21.

[0128]Additional compounds 22-29 were synthesized by the same procedure as
described above (Examples 1 and 2) for Compound 2 except that in Example
1, alternative amines were used in place of L-tert-Leucine methylamide in
the reaction with intermediate Intermediate 1I to form intermediate
Intermediate 1J. For example, Compound 22 was synthesized in the same
manner as Compound 2 except that (S)-2-amino-3,3-dimethylbutan-1-ol was
used in place of L-tert-Leucine methylamide. Compound 23 was synthesized
in the same manner as Compound 2 except that
(R)-2-amino-3,3-dimethylbutan-1-ol was used in place of L-tert-Leucine
methylamide. These resulting intermediates were deprotected with TFA, as
described in Example 2, to form additional Compounds 22-29.

[0134]Compounds 31-38 were synthesized by the same procedure as detailed
above for Compound 30 except that phenyl boronic acid or 4-chlorophenyl
boronic acid was used in step 3 and 1,3,3-trimethylbicyclo
[2.2.1]heptan-2-amine in step 5 was replaced with an alternative amine.
For example, Compound 32 was prepared using aniline as the amine in step
5.

[0140]Compound 49 was synthesized by the same procedure as described above
for Compound 48, above, except that Compound 9 was used in place of
Compound 2. Similarly, Compound 50 was synthesized by the same procedure
as described above for Compound 48 except that Compound 26 was used in
place of Compound 2. In a like manner, Compound 51 was synthesized by the
same procedure as described above for Compound 48 except that compound 27
was used in place of Compound 2.

[0141]Compounds 52-90 were synthesized by the same procedure as described
above for Compound 48, above with the appropriate replacement of Compound
2.

[0144]Compound 92 was synthesized by the same procedure as described above
for Compound 91 except that methylsulfonyl chloride was used in place of
ethanesulfonyl chloride. Similarly, Compound 93 was synthesized by the
same procedure described above for Compound 91 except that
4-fluorobenzenesulfonyl chloride was used in place of ethanesulfonyl
chloride. Likewise, Compound 94 was synthesized by the same procedure as
described above for Compound 91 except that propane-2-sulfonylchloride
was used in place of ethylsulfonylchloride.

[0145]Compounds 95-110 were synthesized by the same procedure as described
above for Compound 91 except for that ethylsulfonylchloride was replaced
with the appropriate sulfonylchloride reagent.

[0148]Compound 112 was synthesized by the same procedure as described
above for Compound 111 except that Compound 39, was used in place of
Compound 45. Compound 113 was synthesized by the same procedure as
described above for Compound 111 except that Compound 40 was used in
place of Compound 45. Compound 114 was synthesized by the same procedure
as described above for Compound 111 except that Compound 41 was used in
place of Compound 45.

[0149]Similarly, compounds 115-118 were synthesized by the same procedure
as described above for compound 111 except that compounds 44, 42, 46 or
47, respectively, were used in place of compound 45.

[0151]To a solution of cyclopropanecarboxylic acid (1.2 eq.) in anhydrous
DMF (2 mL) was distributed EDCI (1.6 eq.), HOBt (1.3 eq.) and TEA (5
eq.). The vial was sealed and stirred for 40 minutes at room temperature.
Compound 39 (1 eq. in 1 mL of anhydrous DMF) was added to the vial which
was then sealed and stirred overnight at room temperature. The solvent
was removed by centrifugal evaporation under reduced pressure. The
residue was dissolved in DCM (2 mL), and washed sequentially with 10%
K2CO3 solution (1 mL) and water (1 mL). The water wash was then
re-extracted with DCM (0.5 mL), the combined organic extracts were
evaporated to dryness under reduced pressure. The desired product,
Compound 119, was isolated by mass directed LC. LCMS (+ESI) m/z 447.38
[M+H]+

[0152]Compounds 120-139 were prepared by the same procedure as described
above for Compound 119 except that another carboxylic acid was used in
place of cyclopropanecarboxylic acid. For example, Compound 122 was
synthesized using furan-2-carboxylic acid.

[0154]To the amine, Compound 45 (1 eq.) in anhydrous DCE (2 mL) was added
TEA (5 eq.). 2-Hydroxyacetaldehyde (1.2 eq. in 1 mL of anhydrous DCE) was
added to the vial, which was sealed and stirred for 1 hour at room
temperature. Sodium triacetoxyborohydride (3 eq.) was added in portions
and the vial was sealed and stirred overnight at room temperature. The
solvent was removed by centrifugal evaporation under reduced pressure.
Saturated sodium carbonate solution (1 mL) was added to the vial, which
was then sealed and sonicated for approximately 20 min DCM (2 mL) was
added and the vial was sonicated for approximately 5 min The organic
layer was removed and the remaining aqueous layer was re-extracted with
DCM (1 mL). The combined organic extracts were evaporated to dryness
under reduced pressure. Compound 140, was purified by mass directed LC.
LCMS (+ESI) m/z 397.18 [M+H]+.

[0155]Compound 141 was synthesized by the same procedure as described
above for Compound 140 except that Compound 47 was used in place of
Compound 45 and tetrahydrofuran-3-carboxaldehyde was used in place of
2-hydroxyacetaldehyde.

[0156]Compounds 142-169 were synthesized by the same procedure as
described above for Compound 140 except that the appropriate reagents
were used in place of Compound 45 and 2-hydroxyacetaldehyde.

[0158]To Intermediate 45 (1 eq. in 1 mL of anhydrous DCM or DMF) was added
TEA (5 eq.). An aliquot of acetyl chloride (1.2 eq. in 1 mL of anhydrous
DCM or DMF) was added to the vial which was then sealed and stirred
overnight at room temperature. The solvent was removed by centrifugal
evaporation under reduced pressure. The residue was dissolved in DCM (2
mL) and washed sequentially with 2M Na2CO3 (1 mL) and water (1
mL). The water wash was then re-extracted with DCM (0.5 mL), the combined
organic extracts and evaporated to dryness under reduced pressure.
Compound 170 was purified by mass directed LC. MS: m/z 395.17
[M+H]+.

[0159]Compound 171 was synthesized by the same procedure as described
above for Compound 170 except that Compound 39 was used in place of
Compound 45 and 2-isocyanatopropane was used in place of acetyl chloride.
Compound 172 was synthesized by the same procedure as described above for
Compound 170 except that Compound 40 was used in place of Compound 45.
Similarly, Compound 173 was synthesized by the same procedure as
described above for Compound 170, except that Compound 41 was used in
place of Compound 45.

[0160]Compounds 174-194 were synthesized by the same procedure as
described above for Compound 170 except that the appropriate reagent was
used in place of Compound 45.

[0164]Step 1: Preparation of intermediate 12B. Intermediate 12A (0.2 g,
0.86 mmol) was dissolved in DMF (3 mL) at 0° C. Dimethylamine (2M
solution in THF, 2.15 mL) was added, followed by HBTU (0.49 g, 1.29
mmol). The reaction mixture slowly warmed to ambient temperature and
stirred overnight. The reaction mixture was concentrated under reduced
pressure and diluted with ethyl acetate. The organic solution was washed
several times with water and dried over anhydrous Na2SO4. The
organic layer was concentrated to provide intermediate 2B which was used
in the next step without further purification. MS: m/z 259.3 [M+H]+.

[0165]Step 2: Preparation of Intermediate 12C. Intermediate 12B was
dissolved in methylene chloride (1 mL) and TFA (1 mL). The reaction
mixture stirred overnight at ambient temperature. The reaction mixture
was concentrated to provide 12C (TFA salt) which was used in the next
step without further purification.

[0171]Compound 198 was synthesized in the same manner as compound 197
except that compound 282 was used in place of compound 2. Similarly,
compound 199 was synthesized in the same manner as compound 197 except
that compound 314 was used in place of compound 2.

[0172]Step 1: Synthesis of (S)-tert-butyl
1-(1-(2-acetylhydrazinyl)-3,3-dimethyl-1-oxobutan-2-ylcarbamoyl)-3-phenyl-
-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate(14D). Intermediate
14C (50 mg, 0.11 mmol) was dissolved in DMF (2 mL) in a 20 mL vial.
Acetyl hydrazide (9 mg, 0.12 mmol) was added followed by DIPEA (38 μL,
0.22 mmol). The mixture was vortexed until homogeneous. TBTU (40 mg, 0.12
mmol) was added and the reaction was stirred for 3 hours, after which
LC/MS showed that the starting material had been consumed. Saturated
NaHCO3 (2 mL) was added to quench and the reaction was extracted of
EtOAc (2×2 mL). The organic layers were combined, dried with
anhydrous Na2SO4, filtered and evaporated to provide
Intermediate 14D as a light yellow oil (48 mg, 85% yield) which was used
in the next step without further purification. LCMS (+ESI) m/z 513.3
[M+H]+.

[0173]Step 2: Synthesis of (S)-tert-butyl
1-(2,2-dimethyl-1-(5-methyl-1,3,4-oxadiazol-2-yl)propylcarbamoyl)-3-pheny-
l-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate (14E). The
intermediate 14D (48 mg, 94 mmol) was taken up in THF (1 mL) and added to
a 2 mL microwave reaction vial. DBU (21 μL, 0.14 mmol) was added to
the reaction, followed by the Burgess reagent (112 mg, 468 mmol). The
vial was capped and the reaction mixture was heated to 150° C. for
5 minutes, after which some starting material still remained. Additional
Burgess reagent (1 equiv) was added and the reaction was heated to
150° C. for 10 minutes. LC/MS showed that the starting material
was consumed. The reaction was diluted with of saturated aqueous
NaHCO3 (1 mL) and EtOAc (2 mL). The organic layer was removed and
the aqueous layer extracted with EtOAc (2 mL). The organic layers were
combined, dried with anhydrous Na2SO4, filtered and evaporated.
The residue was purified by Flash chromatography on silica using a
gradient from 10% EtOAc/Hexanes to 60% EtOAc/Hexanes. This provided
intermediate 14E as a clear oil (25 mg, 54% yield). LCMS (+ESI) m/z 495.3
[M+H]+.

##STR00020##

[0174]Step 3: Synthesis of Compound 200. The intermediate 14E (20 mg, 0.04
mmol) was taken up in 25% TFA in DCM (1 mL). The reaction was stirred for
1 hour after which it was found to be complete by LC/MS. The reaction
mixture was neutralized (pH 7-8). by the addition of saturated aqueous
NaHCO3. The solution was extracted with DCM (3×1 mL). The
combined organic extracts were dried with anhydrous Na2SO4,
filtered and evaporated to provide Compound 200 which was used without
further purification. LCMS (+ESI) m/z 395.0 [M+H]+.

##STR00021##

[0175]Compound 200 was taken up in THF (0.5 mL) and transferred to a 2 mL
microwave vial equipped with a stir bar. Formaldehyde (36 μL, 0.4
mmol) was added followed by the HOAc (3 μL, 0.053 mmol). After mixing,
sodium triacetoxyborohydride (13 mg, 0.061 mmol) was added. The vial was
capped and the reaction was heated at 150° C. for 5 minutes. LC/MS
showed that the reaction was about 70% complete. The reaction was heated
at 160° C. for a further 5 minutes. The reaction was more than 90%
complete by LC/MS, but other products were beginning to form, so the
reaction was terminated. The reaction mixture was diluted with 2 mL of
saturated aqueous NaHCO3 and the solution was vortexed. The solution
was extracted of DCM (3×1 mL). The combined organic extracts were
dried with anhydrous Na2SO4, filtered and evaporated. The
desired product, Compound 201, was obtained as a yellow oil (13 mg, 79%
yield) by flash chromatography on silica using a gradient from 100% EtOAc
to 5% MeOH in EtOAc. LCMS (+ESI) m/z 409.2 [M+H]+.

[0176]Compound 202 was synthesized in the same manner as compound 201
except that isobutryaldehyde was used in place of formaldehyde.

[0178]The intermediate 1J (80 mg, 0.16 mmol) was taken up in 25% TFA in
DCM (2 mL). The reaction was stirred for 1 hour, after which it was found
to be complete by LC/MS. The reaction mixture was neutralized (pH 7-8).
by the addition of saturated aqueous NaHCO3. The solution was
extracted of DCM 3×1 mL. The combined organic layers were dried
with anhydrous Na2SO4, filtered and evaporated. The residue was
dissolved in THF (0.5 mL) and transferred to a 2 mL microwave vial
equipped with a stirrer bar. Formaldehyde (144 μL, 1.6 mmol) was
added, followed by the HOAc (12 μL, 0.21 mmol). After mixing
everything together, the sodium triacetoxyborohydride (50 mg, 0.24 mmol)
was added. The vial was capped and the reaction was heated at 160°
C. for 5 minutes. LC/MS showed that the reaction was more than 90%
complete by LC/MS. The reaction was diluted with saturated aqueous
NaHCO3 (2 mL) and the mixture was vortexed and extracted 3×1
mL of DCM. The combined organic layers were dried with anhydrous
Na2SO4, filtered and evaporated. The desired product, Compound
203, was isolated as a yellow oil (51 mg, 78% yield) by Flash
chromatography on silica using a gradient from 100% EtOAc to 5% MeOH in
EtOAc. LCMS (+ESI) m/z 387.2 [M+H]+.

[0184]Step 2: Synthesis of Intermediate 17B. To a solution of intermediate
17A (0.32 g, 0.70 mmol) in 3:1 THF/water mixture at 0° C. was
added lithium hydroxide (0.84 g, 1.40 mmol). The reaction mixture was
stirred at 0° C. for 6 hours. The reaction was acidified (pH<7)
by dropwise addition of 1N HCl and repeatedly extracted with ethyl
acetate. The combined organic extracts were dried over anhydrous
Na2SO4, filtered and concentrated to provide. Intermediate 17B
(0.3 g) as a crude product, which was used in the next step without
further purification.

[0185]Step 3: Preparation of intermediate 17C. To intermediate 17B (0.3 g,
0.67 mmol) in DMF was added 2M methylamine solution THF (1.7 mL, 3.39
mmol) and TBTU (0.26 g, 0.81 mmol). The reaction mixture stirred for 16
hrs at ambient temperature. The reaction mixture was diluted with water
and extracted several times with ethyl acetate. The combined organic
extracts were dried over anhydrous Na2SO4, filtered and
concentrated under reduced pressure. The residue was purified by flash
chromatography using 0-100% ethyl acetate/hexanes to afford the desired
intermediate 17C (0.1 g).

[0191]Step 3: Preparation of Compound 208. The carboxylic acid
intermediate 18D (400 mg) and DIEA (600 μL) were dissolved in DMF (10
mL). An aliquot of this stock solution (400 μL) was dispensed into a
vial charged with 4-aminobenzene-sulfonamide (0.10 mmol). TBTU was added
(0.50 mmol) to the vial and the mixture was stirred at room temperature
for 2 hours. The crude mixture was purified by prep LC-MS using a
gradient elution of 5% to 95% MeCN/water in 15 min The product was taken
up with DCM and diluted with hexanes. Evaporation under nitrogen flow
gave the desired product, Compound 208 as a solid. LCMS (+ESI) m/z 464
[M+H]+.

[0192]Compounds 209-237 were prepared in the same manner as Compound 208
except that other amines were used in place of
4-aminobenzene-sulfonamide. For example, to synthesize Compound 209,
tert-butyl 4-aminopiperidine-1-carboxylate was used in place of
4-aminobenzenesulfonamide. For compounds 229-237, phenyl boronic was used
in place of 2-fluoro-4-chlorophenylboronic acid in step 1.

[0195]This transformation is also achieved by heating the reaction mixture
in a microwave reactor at 160° C. for 20 minutes. The above
compound is also purified by preparative LC/MS using 5-95% gradient
acetonitrile in water with 0.1% formic acid.

[0196]Compounds 239-273 were synthesized in the same manner as described
for Compound 238 using different boronic acids or pinacol esters in place
of cyclopentenylboronic acid. For example, Compound 254 was synthesized
using 4-cyanophenylboronic acid in place of cyclopentenylboronic acid.

[0197]Compound 274 was synthesized in the same manner as compound 238
except that 3,6-dihydro-2H-pyridine-1-tert-butoxycarbonyl-4-boronic acid
pinacol ester was used in place of cyclopentenylboronic acid and the Boc
group was removed with TFA in DCM using the same procedure as described
in Example 2 above.

[0198]Step 1: Preparation of N-Benzyloxycarbonyl-L-leucine-N-methylamide
(20A). To a solution of N-Benzyloxycarbonyl-L-leucine (1.18 g, 4.45
mmol), methylamine hydrochloride (0.60 g, 8.90 mmol) and DIEA (3.0 mL,
17.2 mmol) in DMF (40 mL) was added TBTU (1.43 g, 5.6 mmol) at 0°
C. in two batches over 10 minutes. After stirring at room temperature
overnight, the reaction was quenched with water (5 mL) and evaporated
under vacuum. The residue was partitioned between brine and EtOAc. The
organic phase was dried over anhydrous Na2SO4, filtered and
evaporated to dryness. The residue was filtered through a short pad of
silica gel with EtOAc to give crude product 20A (1.20 g) which was used
in the next step without purification. LCMS (+ESI) m/z 301.1
[M+Na]+.

[0199]Step 2: Preparation of L-leucine-N-methylamide (20B). A mixture of
intermediate 20A (1.20 g, 4.31 mmol) and 10% palladium on carbon (300 mg)
in MeOH (30 mL) was hydrogenated with a Parr shaker under 55 psi of
hydrogen gas for 3 h. After filtration through celite, the filtrate was
evaporated and azeotroped with EtOAc to give the desired intermediate 20B
as white solid (quant. yield). LCMS (+ESI) m/z 145.1 [M+H]+.

##STR00028##

[0200]Step 3: Preparation of (S)-tert-butyl
3-bromo-1-(4-methyl-1-(methylamino)-1-oxopentan-2-ylcarbamoyl)-5,6-dihydr-
oimidazo[1,5-a]pyrazine-7(8H)-carboxylate (20C). To a solution of
3-bromo-7-(tert-butoxycarbonyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine--
1-carboxylic acid (1I) (0.53 g, 1.53 mmol), L-leucine-N-methylamide (0.26
g, 1.80 mmol) and DIEA (0.5 mL, 2.9 mmol) in DMF (20 mL) was added TBTU
(0.69 g, 2.15 mmol) in two batches over 10 minutes at 0° C. After
stiffing from 0° C. to room temperature for 2 hours, the reaction
was quenched with water and evaporated under vacuum. The residue was
extracted between saturated aqueous NaHCO3 and EtOAc. The organic
layer was dried over anhydrous Na2SO4 and evaporated to
dryness. The crude mixture was purified by column chromatography with 70%
to 100% EtOAc/Hexanes to give the desired product 20C as an oil (48%
yield). LCMS (+ESI) m/z 474.1, 475.1 [M+H]+.

[0202]Step 5: Preparation of
(S)-7-methyl-N-(4-methyl-1-(methylamino)-1-oxopentan-2-yl)-3-phenyl-5,6,7-
,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide (Compound 275). A
mixture of intermediate 20D (50 mg, 0.13 mmol), potassium carbonate (32
mg, 0.23 mmol), phenylboronic acid (0.20 mmol) and palladium tetrakis (8
mg) in dioxane (1.0 mL) and water (0.5 mL) was heated at 100° C.
in a sealed vial overnight. After cooling down to room temperature, the
mixture was passed through a thiol-based palladium scavenger. The residue
was concentrated to dryness, to which MeOH (0.5 mL) was added. The
solution was filtered to remove insoluble material and purified by
preparative LC-MS using a gradient of 5% MeCN/water to 95 MeCN/water
(0.1% formic acid) in 15 min Pure fractions were evaporated with a Savant
speedvac. The resulting oil was taken up in DCM (1.0 mL) and diluted with
hexane (1.0 mL). Evaporation under air flow with mild heating gave
Compound 275 as a white solid product (56% yield). LCMS (+ESI) m/z 384.1
[M+H]+.

[0203]Compounds 276-280 were synthesized in the same manner as Compound
275 except that a different boronic acid was used in place of
phenyboronic acid. For example, for the synthesis of Compound 277,
3-fluoro-4-chlorophenyl boronic acid was used in place of phenylboronic
acid.

[0208]Step 3: Preparation of (S,Z)-tert-butyl
1-(1-(1-aminoethylideneaminooxy)-3,3-dimethyl-1-oxobutan-2-ylcarbamoyl)-3-
-phenyl-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate (Intermediate
22C). Intermediate 22B (0.25 g, 0.55 mmol) was dissolved in DMF and EDCI
(0.17 g, 0.87 mmol) was added, followed by HOBt (0.12 g, 0.87 mmol). The
resulting mixture was stirred for 30 min and (Z)-N'-hydroxy-acetimidamide
(0.06 g, 0.82 mmol) was added and the mixture was stirred overnight. The
reaction mixture was diluted with water and extracted with EtOAc. The
organic layer was washed successively with water, 2N Na2CO3
solution and brine, dried over anhydrous Na2SO4, filtered, and
concentrated under reduced pressure. The residue was used in the next
step without purification. LCMS (+ESI) m/z 1025.2 [2M+H]+.

[0212]Compound 283 was synthesized in the same manner as described above
for Compound 282 except that N'-hydroxy-2,2-dimethylpropanimidamide was
used in place of (Z)-N'-hydroxyacetimidamide in step 3 and the final
compound was methylated using the procedure described in Example 5.

[0213]Compound 284 was synthesized in the same manner as described above
for Compound 282 except that N'-hydroxy-2,2-dimethylpropanimidamide was
used in place of (Z)-N'-hydroxyacetimidamide in step 3 and the final
compound was reacted with (1-ethoxy-cyclopropoxy)trimethylsilane using
the procedure described in Example 13 above.

[0215]Step 1: Preparation of
(S)-3,3-dimethyl-2-(7-methyl-3-phenyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyr-
azine-1-carboxamido)butanoyl chloride (23A). To a solution
(S)-3,3-dimethyl-2-(7-methyl-3-phenyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyr-
azine-1-carboxamido)butanoic acid (Compound 281) (0.11 g, 0.30 mmol) in
DCM was added oxalyl chloride (0.08 g, 0.60 mmol) followed by a catalytic
amount of DMF. The resulting mixture was stirred for 2 h and the solvent
was evaporated, toluene (2 mL) was added and the mixture was concentrated
again. The resulting product (Intermediate 23A) was dried under vacuum
and then used in the next step without further purification.

[0219]Compounds 287 and 288 were synthesized in the same manner as
Compound 286 except that another amine was used in place of morpholine.
For example, Compound 287 was synthesized using piperidine in place of
morpholine.

[0222]Step 2: Preparation of (S)-tert-butyl
3-benzoyl-1-(3,3-dimethyl-1-(methylamino)-1-oxobutan-2-ylcarbamoyl)-5,6-d-
ihydroimidazo[1,5-a]pyrazine-7(8H)-carboxylate (25B). Intermediate 25B was
synthesized in the same manner as described above for compound 22A in
Example 22 except that Intermediate 25A was used in place of 3B and
L-tert-Leucine methylamide was used instead of (S)-methyl
2-amino-3,3-dimethylbutanoate. LCMS (+ESI) m/z 498.3 [M+H]+

[0223]Step 3: Preparation of
(S)-3-benzoyl-N-(3,3-dimethyl-1-(methylamino)-1-oxobutan-2-yl)-5,6,7,8-te-
trahydroimidazo[1,5-a]pyrazine-1-carboxamide (25C). Intermediate 25C was
synthesized in the same manner as described above for Compound 2 except
that Compound 25B was used in place of Compound 1.

[0230]Compound 292 was synthesized by the same procedure as described
above for Compound 48 in Example 5, except that Compound 291 was used in
place of Compound 2. Likewise, compound 293 was synthesized in the same
manner except that in step 2,2-methyl-1-propenylmagnesium bromide was
used in place of neopentyl-magnesium chloride. Similarly, compound 294
was synthesized as described above except that in Step
2,3-phenyl-1-propylmagnesium bromide was used in place of
neopentylmagnesium chloride.

[0234]Compound 203 (20 mg, 52 mmol) was dissolved in 500 μL of THF in a
microwave vial. CuI (10 mg, 52 mmol) and Pd[P(Ph)3]2Cl2
(3.6 mg, 5 μmol) were added followed by the 2-thiazolylzinc (II)
bromide (104 μL of 0.5M solution, 52 umol). The vial was capped and
the mixture was subjected to microwave irradiation with heating to
160° C. for 5 minutes, after which time the reaction was 50%
complete by LC/MS. Addition of further equivalents of zinc reagent
followed by irradiation and heating failed to drive the reaction to
completion.

##STR00037##

[0235]The reaction was quenched by the addition of 2 mL of saturated
aqueous NaHCO3 and extracted with DCM (2×1 mL). The combined
organic layers were dried with anhydrous Na2SO4, filtered and
evaporated. The residue was purified by flash chromatography using a
gradient from 100% EtOAc to 5% MeOH in EtOAc to provide the desired
product, Compound 297, as a yellow solid (2 mg), LCMS (+ESI) m/z 391.3
[M+H]+.

[0237]Compound 2 (15 mg, 35 umol) was dissolved in DMF (3 mL). DIPEA was
added followed by the 2-chloropyrimidine. The reaction was subjected to
microwave irradiation and heated at 160° C. for 15 minutes and
then for a further 5 minutes under the same conditions to drive to
completion. The reaction was quenched by the addition of 2 mL of
saturated aqueous NaHCO3 and extracted of DCM (2×1 mL). The
combined organic layers were dried with anhydrous Na2SO4,
filtered and evaporated. The residue was purified by preparative LC/MS
using a 10 minute gradient from 70% water/acetonitrile to 10%
water/acetonitrile with 0.1% formic acid as a modifier. The desired
product, Compound 298, was isolated as a clear oil (11 mg, 68% yield).
LCMS (+ESI) m/z 448.2 [M+H]+.

[0238]Compounds 299-301 were synthesized in the same manner as described
above for Compound 298 except that different halogenated heterocyles were
used in place of the 2-chloropyrimidine. For example, for Compound 301,
2-chloropyrazine was used in place of 2-chloropyrimidine.

[0241]Step 2: Preparation of (R)-2-amino-N,3,3-trimethylbutanamide.
Intermediate 30A (0.40 g, 1.67 mmol) was dissolved in DCM (3 mL) and TFA
(1.5 mL) was added. The resulting mixture was stirred for 2 hours. The
solvent was evaporated and the residue was diluted with water and
lyophilized to give Intermediate 30B (TFA salt) in a quantitative yield.

[0243]Step 4: Preparation of
(R)--N-(3,3-dimethyl-1-(methylamino)-1-oxobutan-2-yl)-3-phenyl-5,6,7,8-te-
trahydroimidazo[1,5-a]pyrazine-1-carboxamide (Intermediate 30D).
Intermediate 30D was synthesized in the same manner as described above in
Example 2 except that Intermediate 30C was used in place of Compound 1.
LCMS (+ESI) m/z 370.0 [M+H]+.

[0244]Step 5: Preparation of Compound 302. Compound 302 was synthesized in
the same manner as described above in Example 5, except that compound
Intermediate 30D was used in place of Compound 2. LCMS (+ESI) m/z 384.0
[M+H]+.

[0247]Compound 304 was synthesized in the same manner as described above
for compound 303 except compound 243 was used in place of compound 238.
Similarly, compound 305 was synthesized as described except compound 256
was used in place of compound 238. Compound 306 was synthesized in the
same manner except compound 257 was used in place of compound 238.
Likewise, compound 307 was synthesized in the same manner except compound
287 was used in place of compound 238.

[0248]Compound 308 was synthesized in the same manner as described above
for compound 303 except compound 288 was used in place of Compound 238.
Similarly, compound 309 was synthesized as described above except
compound 286 was used in place of compound 238. Compound 310 was
synthesized in a like manner except compound 266 was used in place of
compound 238.

[0252]Step 1: Preparation of Intermediate 33A. Compound 1C (5.4 g) was
stirred with 3-benzylaminopropanol (1.0 equiv) and anhydrous
Na2SO4 (10 g) in THF for 30 min Sodium triacetoxyborohydride
(2.0 equiv) was added and the mixture was stirred at room temperature
over night. The reaction was quenched with brine and extracted with ethyl
acetate. The organic phase was dried over anhydrous Na2SO4 and
evaporated to give crude Intermediate 33A (12.05 g) which was used
without further purification.

[0256]Step 5: Preparation of Intermediate 33E. A solution of Intermediate
33D (2.72 g) and lithium hydroxide monohydrate (0.67 g) in MeOH was
heated at 65° C. for two hours. After evaporation of MeOH, the
residue was extracted between EtOAc and water (pH 3). The aqueous phase
was saturated with sodium chloride and extracted with EtOAc. The combined
organic phase was dried over anhydrous Na2SO4, filtered and
evaporated to give the carboxylic acid Intermediate 33E (2.34 g).

[0257]Step 6: Preparation of Intermediate 33F. To a solution of
Intermediate 33E (0.68 g), L-tert-Leucine-N methylamide (0.37 g) and DIEA
(0.66 mL) in DMF (15 mL) at 0° C. was added TBTU (0.72 g). After
stirring for two hours, the DMF was remove by evaporation under reduced
pressure. The residue was partitioned between saturated aqueous
NaHCO3 and EtOAc. The organic phase was dried over anhydrous
Na2SO4, filtered and evaporated to dryness. The residue was
purified by column chromatography with 80% to 100% EtOAc/hexanes to give
Intermediate 33F (0.52 g).

[0258]Step 7: Preparation of Compound 312. A mixture of Intermediate 33F
(0.52 g), palladium tetrakis(triphenylphosphine) (0.1 g), potassium
carbonate (0.30 g) and phenylboronic acid (0.20 g) in dioxane (10 mL) and
water (5 ml) was heated at 100° C. for two hours. The reaction
mixture was diluted with brine and extracted with EtOAc. The organic
phase was dried over anhydrous Na2SO4, filtered and evaporated
to dryness. The residue was purified by column chromatography with 60% to
100% EtOAc/hexanes to give Compound 312 (0.48 g).

[0259]Compound 313 was prepared in the same manner as Compound 312 except
4-chloro-2-fluorophenyl boronic acid was used in place of phenyl boronic
acid.

[0261]A solution of Compound 312 (0.48 g) in DCM (5 mL) and TFA (5 mL) was
stirred at room temperature for thirty minutes. The solvents were removed
by evaporation under reduced pressure and the residue was extracted
between saturated aqueous NaHCO3 and EtOAc. The aqueous phase was
saturated with sodium chloride and extracted with EtOAc. The combined
organic extracts were dried over anhydrous Na2SO4, filtered and
evaporated to dryness under reduced pressure to give Compound 314 (0.38
g).

[0262]Compound 315 was synthesized in the same manner as described above
for Compound 314 except that 4-chloro-2-fluorophenyl boronic acid was
used in place of phenylboronic acid. Similarly, Compound 316 was
synthesized as described above for Compound 314 except that
4-methyl-2-fluorophenyl boronic acid was used in place of phenylboronic
acid.

[0265]Compound 318 was prepared following the procedure for the synthesis
of Compound 317 except that formaldehyde was replaced with acetaldehyde.
Similarly, Compound 319 was prepared following the same procedure but
replacing formaldehyde with acetone. In a like manner, Compound 320 was
prepared as described for Compound 317, but replacing formaldehyde with
isobutyraldehyde.

[0266]Compound 321 was prepared following the procedure for the synthesis
of compound 317 replacing formaldehyde with cyclopropyl carboxaldehyde.
Compound 322 was prepared following the same procedure, but replacing
formaldehyde with isobutyraldehyde and replacing Compound 314 with
Compound 315. Compound 323 was prepared following the same procedure, but
replacing formaldehyde with cyclopropyl carboxaldehyde and replacing
Compound 314 with Compound 316.

[0268]A solution of Intermediate 33F (0.73 g) in DCM (10 mL) and TFA (10
mL) was stirred at room temperature for one hour. After evaporation of
DCM and TFA, the residue was partitioned between saturated aqueous
NaHCO3 and DCM. The aqueous phase was saturated with sodium chloride
and extracted twice with DCM. The combined organic extracts were dried
over anhydrous Na2SO4, filtered, and evaporated to dryness
under reduced pressure. To a solution of the resulting intermediate (0.55
g), AcOH (82 μL) and 37% aqueous paraformaldehyde (0.5 mL) in THF was
added sodium triacetoxyborohydride (0.60 g). After stiffing at room
temperature overnight, the THF was removed by evaporation under reduced
pressure. The residue was pardoned between brine and DCM and the aqueous
layer was extracted three times with DCM. The combined organic extracts
were dried over anhydrous Na2SO4, filtered and evaporated to
dryness. The residue was purified by column chromatography to give
Compound 324 (387 mg).

[0271]The same reaction was achieved by heating the reaction mixture in a
Biotage microwave reactor at 160° C. for 20 minutes.

[0272]Compounds 326-460, 599-601 were synthesized in the same manner as
compound 325 as described above except other boronic acids or
dioxaborolanes were used in place of 2-methoxyphenylboronic acid. For
example, compound 329 was synthesized using thiophen-3-ylboronic acid in
place of 2-methoxyphenylboronic acid. Compound 366 was synthesized using
4-chloro-2-fluorophenyl boronic acid. Compound 435 was synthesized using
2-cyclopropyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane. Compound 442 was
synthesized using
3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)isoxazole.
Compound 460 was synthesized using
3,6-dihydro-2H-pyridine-1-tert-butoxycarbonyl-4-boronic acid, pinacol
ester and the Boc group was removed with TFA in DCM using the same
procedure as described in Example 34.

[0275]Compound 462 was prepared following the procedure for the synthesis
of compound 461, but replacing ethanesulfonyl chloride with benzoyl
chloride. Similarly, compound 463 was prepared following the procedure
but replacing ethanesulfonyl chloride with acetyl chloride. Likewise,
compound 464 was prepared by following the procedure for the synthesis of
compound 313 except that ethanesulfonyl chloride was replaced with
4-fluorophenylsulfonyl chloride.

[0285]Compound 468 was synthesized in the same manner as described above
for Compound 467 except acetone was used in place of paraformaldehyde.
Similarly, Compound 469 was synthesized in the same manner as described
above for Compound 467 except Compound 466 was used in place of Compound
465. Likewise, compound 470 was synthesized as described above for
Compound 467 except cyclopropane carboxaldehyde was used in place of
paraformaldehyde and Compound 466 was used in place of Compound 465.
Compound 603 was synthesized as described above for Compound 467 except
that compound 602 was used in place of compound 465.

[0290]Compounds 473-480 were synthesized in the same manner as described
above for Compound 472 except that isopropylamine was replaced with
another amine. For example, Compound 473 was synthesized using
n-propylamine in place of isopropylamine.

[0294]Compound 315 (16 mg, 37 umol) was taken up in 3 mL of DMF. DIPEA was
added followed by the 2-chloropyrimidine. The solution was subjected to
microwave irradiation and heated for successive 5 minutes periods, with
increasing temperatures from 125° C. to 160° C. and
addition of more 2-chloropyridine and DIPEA until the reaction was
complete. The reaction was quenched by the addition of saturated aqueous
NaHCO3 (2 mL) and extracted with DCM (2×1 mL). The combined
organic layers were dried with anhydrous Na2SO4, filtered and
evaporated. The residue was purified by automated preparative LC/MS using
a 10 minute method with a gradient from 70% water/acetonitrile to 10%
water/acetonitrile with 0.1% formic acid as a modifier. The desired
product Compound 482 was isolated as a white solid (15 mg, 78% yield).
LCMS (+ESI) m/z 515.1 [M+H]+.

[0295]Compound 483 was synthesized in the same manner as described above
for Compound 482 except that Compound 314 was used in place of Compound
315.

[0296]Compound 434 (42 mg, 0.11 mmol) was dissolved in methanol and was
added to a methanolic slurry of 10% palladium on carbon, Degussa type.
The mixture was subjected to 65 psi of hydrogen gas for 2 hours, filtered
through Celite®, and concentrated under reduced pressure to a yellow
oil.

[0298]Compound 485 was synthesized in the same manner as described above
for compound 484 except compound 376 was used in place of compound 434.
Similarly, compound 486 was synthesized as described above for compound
484 except compound 377 was used in place of compound 434. Likewise,
compound 487 was synthesized in the same manner as described above for
compound 484 except compound 396 was used in place of compound 434.
Compounds 488-506 were synthesized in the same manner as described above
for compound 484 except compound 434 was replaced with the appropriate
reagent.

[0299]Compound 507 was synthesized in the same manner as described above
for compound 484 except compound 454 was used in place of Compound 434.
Compound 508 was similarly synthesized as described above except that
compound 514 was used in place of compound 434.

[0301]Compound 510 was synthesized in the same manner as compound 509
except piperidine was used in place of morpholine. Similarly, compound
511 was synthesized as described for compound 509 except pyrrolidine was
used in place of morpholine. Compound 512 was synthesized in the same
manner except diethylamine was used in place of morpholine. Compound 513
was synthesized in the same manner except that dimethylamine was used in
place of morpholine.

[0304]Preparation of
(S)-2-(8-(tert-butoxycarbonyl)-3-phenyl-6,7,8,9-tetrahydro-5H-imidazo[1,5-
-a][1,4]diazepine-1-carboxamido)-3,3-dimethylbutanoic acid (Compound 515).
To a solution of Intermediate 39C (571 mg, 0.77 mmol) in tetrahydrofuran
(2 mL) were added a 10M aqueous solution of sodium hydroxide (0.77 mL,
7.7 mmol) and methanol (2 mL). The solution was stirred for 2.5 hours and
was then concentrated under reduced pressure. The residue was brought to
pH 3 with 1N aqueous hydrochloric acid solution, and a resulting
precipitate was collected by filtration and washed with water.

[0315]Compound 522 was prepared in the same manner as described above for
compound 521 except that compound 520 was used in place of compound 519.
Similarly, compound 523 was prepared in the same manner except that
compound 520 was used in place of compound 519 and acetone was used in
place of formaldehyde. Likewise, compound 524 was prepared as described
above for compound 521 except that acetaldehyde was used in place of
formaldehyde. Compound 525 was prepared in the same manner except that
acetone was used in place of formaldehyde.

[0320]Step 1: A mixture of compound 448 (0.65 g, 1.41 mmol) and palladium
on carbon (380 mg) in methanol was hydrogenated under 60 psi hydrogen for
2 hours. After filtration of catalyst, the solution was evaporated to
dryness. The residue (0.54 g, 1.17 mmol) was stirred with acetone and 2N
HCl at room temperature overnight. After evaporation of acetone, the
aqueous phase was basified with saturated aqueous NaHCO3 and
extracted with 10% iPrOH/DCM twice. The combined organic phase was dried
over anhydrous Na2SO4 and evaporated to dryness to give the
ketone intermediate 54A at 92% yield. LCMS (+ESI) m/z 418.3, [M+H]+.

[0321]Step 2: To a solution of intermediate 54A (0.45 g, 1.08 mmol) in DCE
was added DAST (0.40 g, 2.47 mmol) and then heated at 80° C. for 2
hours. The reaction was quenched with aqueous NaHCO3 and extracted
with 10% iPrOH/DCM twice. The combined organic phase was dried over
anhydrous Na2SO4 and evaporated to dryness. The residue was
purified by reverse phase column chromatography with 20% to 60%
MeCN/water (0.5 formic acid). Fractions with product were combined and
lyophilized to dryness. The obtained solid was dissolved in MeOH and
purified by preparative LC-MS with 5% to 95% MeCN/water to give compound
528 at 2% yield. LCMS (+ESI) m/z 440.4, [M+H]+.

[0326]Compounds 530-539 were synthesized in the same manner as described
above for compound 529 except that 4-chloro-2-fluorophenyl boronic acid
was replaced with another boronic acid or dioxaborolane. For example,
compound 530 was synthesized in the same manner as compound 529 except
that 4-fluorophenyl boronic acid was used in place of
4-chloro-2-fluorophenyl boronic acid.

[0327]Compound 540 was synthesized in the same manner as described above
for compound 529 except that 4-chloro-2-fluorophenyl boronic acid was
replaced with cyclohexene-1-boronic acid, pinacol ester. The cyclohexenyl
intermediate was then reduced using the hydrogenation procedure described
in Example 45.

[0328]Compounds 541 and 542 were synthesized in the same manner as
described above for compound 529 except that
(S)-2-amino-2-cyclohexyl-N-methylacetamide (synthesized in the same
manner as Intermediate 20B, Example 20) was used in place of 20B and
4-chloro-2-fluorophenyl boronic acid was replaced with
2,4,5-trifluorophenyl boronic acid or 2,4-difluoro-5-chlorophenylboronic
acid pinacol ester.

[0330]Step 1: To a solution of carboxylic acid 33E (100 mg, 0.28 mmol),
phenyl glycine methylamide (56A) (86 mg, 0.53 mmol, prepared according to
the procedure for the synthesis of leucine-N-methylamide by replacing
Z-Leu-OH with Z-Phg-OH in Example 20 and DIEA (100 μL, 0.58 mmol) in
DMF was added TBTU (134 mg, 0.53 mmol) at 0° C. After stirring at
from 0° C. to room temperature for 4 h, the reaction was quenched
with water (5 mL) and evaporated under vacuum. The residue was extracted
between brine and EtOAc. The organic phase was dried over anhydrous
Na2SO4 and evaporated to dryness. The residue was purified by
column chromatography with 70% to 100% EtOAc/Hex to give Intermediate 56B
at 56% yield. LCMS (+ESI) m/z 508.0 [M+Na]+.

[0331]Step 2: A mixture of 56B (100 mg, 0.20 mmol), phenylboronic acid (36
mg, 0.30 mmol), potassium carbonate (46 mg, 0.33 mmol) and palladium
tetrakis(triphenylphosphine) (10 mg) in dioxane and water (3:1) was
heated at 100° C. overnight. The reaction mixture was diluted with
brine and extracted with EtOAc. The organic phase was dried and
evaporated to dryness and purified by column chromatography with 75% to
100% EtOAc/Hexanes to give Intermediate 56C at 54% yield.

[0336]Compounds 545-555 were synthesized in the same manner as described
above for compound 544 except that 4-chloro-2-fluorophenyl boronic acid
was replaced with another boronic acid or dioxaborolane. For example,
compound 547 was synthesized in the same manner as compound 544 except
that 4-fluorophenyl boronic acid was used in place of
4-chloro-2-fluorophenyl boronic acid.

[0340]Compounds 557-563 were synthesized in the same manner as described
above for compound 556 except that 4-chloro-2-fluorophenyl boronic acid
was replaced with another boronic acid or dioxaborolane. For example,
compound 561 was synthesized in the same manner as Compound 556 except
that 3,4-difluorophenyl boronic acid was used in place of
4-chloro-2-fluorophenyl boronic acid.

[0344]Compounds 565-572 were synthesized in the same manner as described
above for Compound 564 except that 4-chloro-2-fluorophenyl boronic acid
was replaced with another boronic acid or dioxaborolane. For example,
Compound 565 was synthesized in the same manner as Compound 564 except
that 4-difluorophenyl boronic acid was used in place of
4-chloro-2-fluorophenyl boronic acid. Compound 570 was synthesized in the
same manner as described above for Compound 564 except that
4-chloro-2-fluorophenyl boronic acid was replaced with
cyclohexene-1-boronic acid, pinacol ester. The cyclohexenyl intermediate
was then reduced using the hydrogenation procedure described in Example
45.

[0348]Compounds 574-582 were synthesized in the same manner as described
above for compound 573 except that 4-chloro-2-fluorophenyl boronic acid
was replaced with another boronic acid or dioxaborolane. For example,
Compound 582 was synthesized in the same manner as Compound 573 except
that phenyl boronic acid was used in place of 4-chloro-2-fluorophenyl
boronic acid.

[0352]Compounds 584-591 were synthesized in the same manner as described
above for Compound 583 except that 2,5-difluorophenyl boronic acid was
replaced with another boronic acid or dioxaborolane. For example,
compound 584 was synthesized in the same manner as 583 except that
2-fluoro-4-chloro phenyl boronic acid was used in place of
2,5-difluorophenyl boronic acid.

[0358]Compound 593 was synthesized in the same manner as described above
for compound 592 except that 2,4,5-trifluorophenyl boronic acid was
replaced with 2,4-difluoro-5-chlorophenylboronic acid pinacol ester.

[0360]Step 1. Preparation of 3,6-dihydro-2H-pyran-4-yl
trifluoromethanesulfonate (Intermediate 63A). Dihydro-2H-pyran-4(3H)-one
(0.3 g, 3.30 mmol) was dissolved in THF and
1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide
(1.18 g, 3.30 mmol) was added. The resulting mixture was cooled to
-78° C. and lithium bis(trimethylsilyl)amide (1M in THF, 3.30 mL)
was added dropwise and the reaction mixture was stirred at -78° C.
for 2 hours. The mixture was then allowed to warm to -5° C. over a
period of 15 hours. The reaction mixture was quenched with saturated
ammonium chloride solution and extracted with ethyl acetate. The organic
extracts were washed with brine, dried over Na2SO4 and
concentrated. The residue was purified by regular phase chromatography
eluting with 10% ethyl acetate/hexanes. The resulting crude oil 63A (200
mg) was used in the next step without purification.

[0361]Step 2. Preparation of
2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(intermediate 63B). Intermediate 63A (0.20 g 0.86 mmol) was dissolved in
dioxane and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane)
(0.32 g, 1.29 mmol) was added followed by KOAc (0.25 g, 1.29 mmol). The
resulting mixture was degassed with nitrogen and PdCl2(dPPF) (0.050
g, 0.069 mmol) was added. The resulting mixture was heated at 80°
C. overnight. The mixture was concentrated, and the oily residue was
extracted with ethyl acetate. The organic extracts were concentrated and
the residue was purified by regular phase chromatography eluting with 10%
ethylacetate/hexanes to give a title compound 63B as a clear oil which
was used in the next step without purification.

[0363]Step 1. Preparation of ethyl
3-phenyl-6,7,8,9-tetrahydro-5H-imidazo[1,5-a][1,4]diazepine-1-carboxylate
(64A). Intermediate 39A (1.22 g, 3.17 mmol) was dissolved in DCM (10 mL)
and TFA (5 mL) was added. The resulting mixture was stirred for 2 hours.
The mixture was concentrated, toluene was added and the mixture was
concentrated again and dried under vacuum to give 1.28 g of intermediate
64A which was used in the next step without purification. LCMS (+ESI) m/z
286.1 [M+H]+

[0367]Step 4. Preparation of
8-methyl-3-phenyl-6,7,8,9-tetrahydro-5H-imidazo[1,5-a][1,4]diazepine-1-ca-
rbonyl chloride (64D). To a solution of intermediate 64C (0.12 g, 0.44
mmol) in 3 mL DCM was added thionyl chloride (0.16 mL, 2.21 mmol),
followed by catalytic amount of DMF. The resulting mixture was stirred at
50° C. for 30 minutes, concentrated and the residual thionyl
chloride was azeotroped with toluene. The resulting material 64D was
dried under vacuum and used in the next step without purification.

[0385]Disclosure of the structures and detailed synthesis and properties
of a further three hundred substituted imidazoheterocycles having the
structure of formula I can be found in the related application, U.S. Ser.
No. 12/337,683 published as US Patent Application Publication No. US
2009/0149450 A1, the specification of which is hereby incorporated by
reference.

[0389]On the day of the experiment, cells were spun at low speed for 5 min
at room temperature. The supernatant was removed and cells were
resuspended in stimulation buffer (Hanks Buffered Salt Solution/5 mM
HEPES, containing 0.5 mM IBMX (cat #17018, Sigma) and 0.02% BSA
(Perkin-Elmer, cat #CR84-100)). Cell clumps were removed by filtering
through cell strainer 40 μm (BD Falcon, Discovery Labware, Bedford,
Mass.) and diluted to 2×105 cells/mL. Antibody supplied with
the LANCE cAMP immunoassay kit was then added according to the
manufacturer's instructions. An aliquot of cells was taken for un-induced
controls. To the remaining cells was added NKH-477 (a water soluble
forskolin derivative, Tocris cat #1603) to a final concentration of 2-8
μM. Cells were then incubated for 30 min at room temperature prior to
adding to Proxiplates containing test compounds (final DMSO concentration
was less than 0.5%) with a Multidrop bulk dispenser, followed by a sixty
minute incubation at room temperature. The response was stopped by
addition of the detection mix supplied with the LANCE kit. The reagents
were allowed to equilibrate for three hours prior to reading on an
Envision multi-mode detector (Perkin-Elmer). TR-FRET was measured using a
330-380 nm excitation filter, a 665 nm emission filter, dichroic mirror
380 nm and Z=1 mm Cyclic AMP concentrations in each well were
back-calculated from a cAMP standard curve run concurrently during each
assay. Each plate contained 16 wells of forskolin stimulated cells and 16
wells of forskolin plus CP55,940-treated cells. Cells were treated with 1
μM CP55,940 (Tocris cat. # 0949). Concentrations of cAMP were
expressed as a percent of the difference of these two groups of wells.
Concentration-response data including EC50 (the concentration of
compound producing 50% of the maximal response) and intrinsic activity
(the percent maximal activation compared to full activation by CP55,940)
were determined using a 4-parameter non-linear regression algorithm
(Xlfit equatn 251, IDBS).

Example 85

Determination of EC50 values for Compounds at Human and Rat
Cannabinoid Receptors

[0390]Tables IIA, IIB and IIC, below show compounds (1)-(607) and Tables
IIIA and IIIB show compounds (608)-(914) grouped by EC50 ranges. For
convenience, the ranges were chosen as follows: The most potent group of
compounds was classified in an EC50 range from 0.1 nM to 10 nM. The
second most potent group was classified in an EC50 range from
greater than 10 nM to 100 nM. The third most potent group was classified
in an EC50 range from greater than 100 nM to 10 μM. Finally, the
fourth group classified by potency had an EC50 of greater than 10
μM determined as above.

[0391]The EC50 range for each of the compounds 1-607 determined
against the human CB2 receptor (hCB2 EC50) is shown in Table IIA.

[0392]Table IIIA shows the hCB2 EC50 ranges for compounds 608-914.

[0393]The EC50 range for each of the compounds 1-607 determined
against the rat CB2 receptor is shown in Table IIB.

[0394]The EC50 range for each of the compounds 1-607 determined
against the human CB1 receptor (hCB1 EC50) is shown in Table IIC.

[0396]The anti-hyperalgesic effects of test compounds in the Complete
Freund's Adjuvant (CFA) model of inflammatory pain was examined as
described below. Male Sprague-Dawley rats (Hsd:Sprague-Dawley®®
SD®®, Harlan, Indianapolis, Ind.) weighing 201±1 grams, were
housed three per cage Animals had free access to food and water and were
maintained on a twelve hour light/dark schedule for the entire duration
of the experiment. Approximately 12 hours prior to behavioral testing,
animals were placed on wire mesh bottom cages with free access to water
but no access to food. Test compounds were prepared in 50% PEG-400
(Sigma-Aldrich, cat. P3265). Indomethacin (Fluka, cat 57413) was
suspended in 0.5% methylcellulose (Sigma-Aldrich, cat. 274429). Groups of
eight animals were anesthetized with 2-3% isoflurane and local
inflammation induced by 50 μl CFA (Sigma-Aldrich, cat F5881,
Mycobacterium tuberculosis 1 mg/ml) injected subcutaneously into the
plantar surface of the left paw.

[0397]Assessment of mechanical hyperalgesia: Baseline and post-treatment
withdrawal thresholds to a noxious mechanical stimulus were measured
using the Randall-Selitto paw pressure apparatus (Ugo Basile
Analgesymeter, model 7200). This apparatus generates a linearly
increasing mechanical force. The stimulus was applied to the plantar
surface of the hind paws by a dome-shaped plastic tip placed between the
third and fourth metatarsus. To avoid tissue damage, a cut-off pressure
was set at 390 grams. Mechanical thresholds were defined as the force in
grams at the first pain behavior, which includes paw withdrawal,
struggle, and/or vocalization. Indomethacin (30 mg/kg, p.o.) served as
the positive control. Mechanical hyperalgesia was measured using the
Randall-Selitto paw pressure device before CFA injection and after
intraperitoneal (i.p.) compound administration over a twenty-four-hour
period. The mean and standard error of the mean (SEM) were determined for
the injured and normal paws for each treatment group. The results for
Compound 91 as compared with vehicle alone are shown in FIG. 1. No side
effects were observed during the course of the experiment.

Example 87

Inhibition of Acetic Acid-Induced Writhing in Mice

[0398]This test identifies compounds which exhibit analgesic activity
against visceral pain or pain associated with activation of low
pH-sensitive nociceptors [see Barber and Gottschlich (1986) Med. Res.
Rev. 12: 525-562; Ramabadran and Bansinath (1986) Pharm. Res. 3:
263-270]. Intraperitoneal administration of dilute acetic acid solution
causes a writhing behavior in mice. A writhe is defined as a contraction
of the abdominal muscles accompanied by an extension of the forelimbs and
elongation of the body. The number of writhes observed in the presence
and absence of test compounds is counted to determine the analgesic
activity of the compounds.

[0399]Male ICR mice, 20-40 grams in weight, were weighed and placed in
individual observation chambers (usually a 4000 ml beaker) with a fine
layer of rodent bedding at the bottom. To determine the activity and
potency of test compounds, different doses of the compound solution or
vehicle were injected subcutaneously in the back of the neck 30 minutes
prior to administration of acetic acid solution. After administration of
the compound or vehicle control, mice were returned to their individual
observation chambers awaiting the intraperitoneal administration of
acetic acid solution. Thirty minutes later, 10 ml/kg of a 0.6% (v/v)
acetic acid solution was then injected into the right lower quadrant of
the abdomen. Immediately after the injection, the mouse was returned to
its observation chamber and the recording of the number of writhes begun
immediately. The number of writhes was counted over a 15-min period
starting from the time of acetic acid injection. Raw data were analyzed
using a one-way ANOVA followed by Dunnett's post-tests. For dose-response
analysis, raw data were converted to % maximum possible effect (% MPE)
using the formula: % MPE=((Wc-Wv)/(0-Wv))*100, where Wc is the number of
writhes in compound-treated mice and Wv is the mean number of writhes in
vehicle-treated mice. The dose which elicited 50% attenuation of
hypersensitivity (ED50) was determined using linear regression analysis.
(Tallarida & Murray, 1987).

[0400]Dose response relationships were established for compounds 317 and
366 by subcutaneous injection of doses equivalent to 3, 10 and 30 mg/kg
given 30 minutes before intraperitoneal injection of the acetic acid
solution. The number of writhes observed in treated an untreated animals
were compared. Results are shown in FIG. 2.

Example 88

Carrageenan Model of Acute Inflammation

[0401]Acute inflammation was produced in rats by injecting 0.1 mL of 2%
λ-carrageenan (type IV; Sigma, St. Louis, Mo.) into one hind paw.
Carrageenan treatment elicited a marked hind paw swelling (edema)
relative to the non-injected paw. At various time points following
carrageenan injection, paw volume measurements were taken for both hind
paws using a plethysmometer (Stoelting). Briefly, the rat was gently held
under the arms with one hand, and its ankle was stabilized with the other
hand, each paw was dipped (for a duration of ˜sec, i.e., sufficient
time to get a stable reading) into a known volume of fluid and total
fluid displacement was recorded Animals were administered vehicle or test
compounds prior to carrageenan administration. A statistically
significant reduction in hind paw volume relative to the vehicle-treated
control group was interpreted as an anti-inflammatory effect.

[0403]The SNL model (Kim and Chung 1992) was used to induce chronic
neuropathic pain in rats. The rats were anesthetized with isoflurane, the
left L5 transverse process was removed, and the L5 and L6 spinal nerves
were tightly ligated with 6-0 silk suture. The wound was then closed with
internal sutures and external staples. Following at least seven days post
SNL, baseline, post-injury and post-treatment values for non-noxious
mechanical sensitivity were evaluated using eight Semmes-Weinstein
filaments (Stoelting, Wood Dale, Ill., USA) with varying stiffness (0.4,
0.7, 1.2, 2.0, 3.6, 5.5, 8.5, and 15 g) according to the up-down method
(Chaplan et al. 1994). Animals were placed on a perforated metallic
platform and allowed to acclimate to their surroundings for a minimum of
thirty minutes before testing. The mean and standard error of the mean
(SEM) were determined for the injured paw in each treatment group. Since
this stimulus is normally not considered painful, significant
injury-induced increases in responsiveness in this test are interpreted
as a measure of mechanical allodynia. The dose which elicited 50%
attenuation of mechanical hypersensitivity (ED50) was determined
using linear regression analysis. Results obtained after oral
administration of compound 317 at 3, 10 and 30 mg/kg and with compound
366 at 1, 3 and 10 mg/kg are shown in FIG. 4.

Example 90

Cytokine Production by Human Macrophages

[0404]Cytokine production by LPS-induced monocytes is a model system for
testing anti-inflammatory candidate molecules. Cytokines induced by LPS
include TNFα, IL-1β, IL-6, and IL-8. Induction of these
cytokines is increased by interferon-γ (IFNγ).

[0406]Cytokines were assayed by a bead-based assay on a Luminex 100
according to the manufacturer's instructions. Statistical comparison was
made by one way ANOVA combined with post-analysis Dunnet's Multiple
Comparison test using Graphpad Prism version 5 software. Statistical
significance was accepted at P<0.05. Results are shown in Table III
below.

[0408]The texts of the references cited in this specification are herein
incorporated by reference in their entireties. In the event that a
definition of a term as incorporated by reference differs from the
meaning defined herein, then the meaning provided herein is intended. The
examples provided herein are for illustration purposes only and are not
to be interpreted as limiting the scope of the invention, the full scope
of which will be immediately recognized by those of skill in the art.